what are new research topics in forensic science

299+ Forensic Science Research Topics (Updated 2024)

Welcome to the world of Forensic Science Research Topics. Get ready to dive into a treasure of fascinating ideas that crack the mysteries behind crime-solving techniques. This year’s collection spans 15 stunning categories, each including 20 engaging topics.

From DNA Analysis uncovering secrets in genes to Ballistics & Firearms exploring the science behind bullets, these categories open doors to understanding how science solves puzzling cases. Cyber Forensics delves into the digital world of crime, while Forensic Anthropology examines the stories hidden within skeletal remains.

Explore Toxicology & Drug Analysis, diving into the science of poisons and medications, or journey into Wildlife Forensics, where nature meets investigation. Uncover the secrets of Bloodstain Pattern Analysis or delve into the linguistic clues in Forensic Linguistics.

Join us as we uncover the mysteries, piece by piece, and go on a thrilling journey into the captivating realm of Forensic Science Research for the year 2024.

Top 5 Applications of Forensic Science

Table of Contents

Forensic science stands as a crucial pillar in solving mysteries within the kingdoms of crime and justice. It includes various scientific disciplines applied to legal matters, providing key insights that aid investigations and legal proceedings. This multidisciplinary field plays a crucial role in solving crimes, identifying culprits, and bringing closure to victims’ families. Here are the top 5 applications of Forensic Science:

Crime Scene Investigation (CSI) : Forensic science’s foundation involves detailed examination of crime scenes. It encompasses evidence collection, analysis of fingerprints, bloodstains, fibers, and other trace evidence. This critical process helps reconstruct the sequence of events leading to a crime.
DNA Analysis : The advancement in DNA technology has revolutionized forensic science. DNA analysis helps identify individuals, link suspects to crime scenes, and exonerate innocent parties. It’s a powerful tool in criminal investigations and solving cold cases.
Toxicology & Drug Analysis : Forensic toxicology focuses on detecting drugs, poisons, or toxins in the body. It’s instrumental in determining causes of death or establishing impairment due to substances.
Ballistics & Firearms Analysis : This branch involves studying firearms, bullets, and cartridge cases. It assists in linking weapons to crimes, identifying shooting distances, and determining trajectories.
Digital Evidence Examination : In the digital age, forensic science extends into cyberspace. Experts analyze digital devices and data to recover, interpret, and present evidence pertinent to cybercrimes.

These five applications showcase how forensic science’s diverse toolkit and methodologies are instrumental in solving crimes, offering justice, and ensuring a safer society.

Top 299+ Forensic Science Research Topics

Now, join us in exploring these thought-provoking themes and be part of the thrilling journey where every clue leads to a new revelation in the field of forensic research. Let us start.

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Recent Dissertation Topics in Forensic Science

This article serves as a compass, guiding readers through a diverse array of recent dissertation topics that encapsulate the multifaceted nature of forensic research. From digital forensics to forensic psychology, the chosen dissertation topics reflect the evolving challenges and advancements in solving complex legal puzzles.

Forensic DNA Analysis:

“Next-Generation Sequencing (NGS) in Forensic DNA Profiling: Opportunities and Challenges”
“The Impact of DNA Transfer and Secondary DNA Transfer in Forensic Investigations”
“Ethical Implications of DNA Phenotyping: A Critical Analysis”

Digital Forensics:

“Artificial Intelligence in Digital Forensic Analysis: A Comprehensive Review”
“Cloud Forensics: Investigating Digital Crimes in Cloud Computing Environments”
“Deepfake Detection Techniques: Safeguarding Digital Evidence Integrity”

Forensic Anthropology:

“Facial Approximation in Forensic Anthropology: Integrating 3D Modeling Techniques”
“The Role of Forensic Anthropologists in Mass Graves Investigations”
“Advancements in Skeletal Trauma Analysis for Forensic Purposes”

Forensic Toxicology:

“Metabolomics in Forensic Toxicology: Profiling Endogenous and Exogenous Compounds”
“Designer Drugs: Analytical Approaches for the Detection of Novel Psychoactive Substances”
“Forensic Challenges in Analyzing Postmortem Fluids for Toxicological Investigations”

Forensic Psychology:

“The Impact of Jury Bias on Forensic Psychologists’ Testimonies: A Case Study Analysis”
“Virtual Reality Applications in Forensic Psychology Training: Enhancing Investigative Skills”
“Exploring the Ethical Dilemmas in Forensic Psychological Assessments”

Forensic Pathology:

“Cardiac Biomarkers in Forensic Pathology: Exploring their Role in Cause of Death Determination”
“The Use of Postmortem Imaging in Forensic Pathology: A Comparative Analysis”
“Forensic Aspects of Pediatric Traumatic Brain Injuries: Patterns and Challenges”

Forensic Odontology:

“Age Estimation in Subadults: Integrating Dental and Skeletal Methods in Forensic Odontology”
“Digital Methods in Bite Mark Analysis: Enhancing Accuracy and Reliability”
“Role of Dental Records in Disaster Victim Identification: A Global Perspective”

Forensic Entomology:

“Forensic Entomogenomics: Unraveling New Dimensions in Time of Death Estimation”
“Environmental Factors Influencing Insect Colonization on Decomposing Remains: A Forensic Study”
“The Use of Entomotoxicology in Forensic Investigations: Current Trends and Applications”

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January 7, 2024 at 2:22 am

thank you for this post. I needed to submit a topic for my dissertation on Monday and you guys saved me big time

Recent advances in forensic science research

For immediate release, acs news service weekly presspac: april 20, 2022.

Forensic scientists collect and analyze evidence during a criminal investigation to identify victims, determine the cause of death and figure out “who done it.” Below are some recent papers published in ACS journals reporting on new advances that could help forensic scientists solve crimes. Reporters can request free access to these papers by emailing newsroom@acs.org .

“Insights into the Differential Preservation of Bone Proteomes in Inhumed and Entombed Cadavers from Italian Forensic Caseworks” Journal of Proteome Research March 22, 2022 Bone proteins can help determine how long ago a person died (post-mortem interval, PMI) and how old they were at the time of their death (age at death, AAD), but the levels of these proteins could vary with burial conditions. By comparing bone proteomes of exhumed individuals who had been entombed in mausoleums or buried in the ground, the researchers found several proteins whose levels were not affected by the burial environment, which they say could help with AAD or PMI estimation.

“Carbon Dot Powders with Cross-Linking-Based Long-Wavelength Emission for Multicolor Imaging of Latent Fingerprints” ACS Applied Nanomaterials Jan. 21, 2022 For decades, criminal investigators have recognized the importance of analyzing latent fingerprints left at crime scenes to help identify a perpetrator, but current methods to make these prints visible have limitations, including low contrast, low sensitivity and high toxicity. These researchers devised a simple way to make fluorescent carbon dot powders that can be applied to latent fingerprints, making them fluoresce under UV light with red, orange and yellow colors.

“Proteomics Offers New Clues for Forensic Investigations” ACS Central Science Oct. 18, 2021 This review article describes how forensic scientists are now turning their attention to proteins in bone, blood or other biological samples, which can sometimes answer questions that DNA can’t. For example, unlike DNA, a person’s complement of proteins (or proteome) changes over time, providing important clues about when a person died and their age at death.

“Integrating the MasSpec Pen with Sub-Atmospheric Pressure Chemical Ionization for Rapid Chemical Analysis and Forensic Applications” Analytical Chemistry May 19, 2021 These researchers previously developed a “MasSpec Pen,” a handheld device integrated with a mass spectrometer for direct analysis and molecular profiling of biological samples. In this article, they develop a new version that can quickly and easily detect and measure compounds, including cocaine, oxycodone and explosives, which can be important in forensics investigations.

The American Chemical Society (ACS) is a nonprofit organization chartered by the U.S. Congress. ACS’ mission is to advance the broader chemistry enterprise and its practitioners for the benefit of Earth and all its people. The Society is a global leader in promoting excellence in science education and providing access to chemistry-related information and research through its multiple research solutions, peer-reviewed journals, scientific conferences, eBooks and weekly news periodical Chemical & Engineering News . ACS journals are among the most cited, most trusted and most read within the scientific literature; however, ACS itself does not conduct chemical research. As a leader in scientific information solutions, its CAS division partners with global innovators to accelerate breakthroughs by curating, connecting and analyzing the world’s scientific knowledge. ACS’ main offices are in Washington, D.C., and Columbus, Ohio.

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Articles on Forensic science

Displaying 1 - 20 of 117 articles.

what are new research topics in forensic science

Your fingerprint is actually 3D − research into holograms could improve forensic fingerprint analysis

Partha Banerjee , University of Dayton

The strange story of the grave of Copernicus

Darius von Guttner Sporzynski , Australian Catholic University

The words that helped wrongly convict Kathleen Folbigg

Kate Burridge , Monash University and Stephen Cordner , Victorian Institute of Forensic Medicine

Juries that don’t understand forensic science can send innocent people to prison − a short training video could help

Devon LaBat , Florida International University ; Deborah Goldfarb , Florida International University ; Jacqueline R. Evans , Florida International University , and Nadja Schreiber Compo , Florida International University

Your unique body odor could identify who you are and provide insights into your health – all from the touch of a hand

Chantrell Frazier , Framingham State University ; Kenneth G. Furton , Florida International University , and Vidia A. Gokool , Lawrence Livermore National Laboratory

New research reveals that Ötzi the iceman was bald and probably from a farming family – what else can DNA uncover?

Caroline Smith , University of Westminster

Secrets wrapped in fabric: how our study of 100 decomposing piglet bodies will help solve criminal cases

Paola A. Magni , Murdoch University and Ian Dadour , Murdoch University

The Titan disaster investigation has begun. An expert explains what might happen next

Paola A. Magni , Murdoch University

Kathleen Folbigg pardon shows Australia needs a dedicated body to investigate wrongful convictions

David Hamer , University of Sydney and Andrew Dyer , University of Sydney

Who owned this Stone Age jewellery? New forensic tools offer an unprecedented answer

Michelle Langley , Griffith University

The camera never lies? Our research found CCTV isn’t always dependable when it comes to murder investigations

Helen Jones , University of South Wales and Fiona Brookman , University of South Wales

Lie detection tests have worked the same way for 3,000 years – and they’re still hopelessly inaccurate

Rebecca Wilcoxson , CQUniversity Australia and Emma Turley , CQUniversity Australia

Six parts of your car that gather data on you

Rachael Medhurst , University of South Wales

DNA is often used in solving crimes. But how does DNA profiling actually work?

Adrian Linacre , Flinders University

Children’s eyewitness testimony can be as accurate as adults’ or more so – if interviewers follow these guidelines

Ben Cotterill , Clemson University

When remains are found in a suitcase, forensics can learn a lot from the insects trapped within

How Nairobi police failures let people get away with murder

Douglas Lucas Kivoi , The Kenya Institute for Public Policy Research and Analysis (KIPPRA)

Identifying the dead after mass disasters is a crucial part of grieving. Here’s how forensic experts do it

Flies, maggots and methamphetamine: how insects can reveal drugs and poisons at crime scenes

How centuries-old bones from Australia’s historic shipwrecks can help us solve crimes

Paola A. Magni , Murdoch University and Edda Guareschi , Murdoch University

Top contributors

Associate professor of Forensic Science, Murdoch University

Visiting Scholar, University of Wollongong in Dubai (UOWD), University of Wollongong

Adjunct Associate Professor, University of New England

Adjunct professor, Murdoch University

Research fellow, The University of Queensland

Chancellor’s Postdoctoral Research Fellow, University of Technology Sydney

Professor of Forensic Science, University of Hull

Forensic Pathologist at Monash University and Deputy Director, Victorian Institute of Forensic Medicine

Reader, School of Natural and Built Environment, Queen's University Belfast

Lecturer in Chemistry, University of Surrey

Assistant Professor, Northumbria University, Newcastle

Associate Professor of Archaeological Science, Flinders University

Associate Dean for Academic Affairs and Professor of Law, Georgia State University

Reader in Forensic Geoscience, Keele University

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Big footsteps and new challenges

Open access
Published: 03 May 2022
Volume 18 , pages 123–124, ( 2022 )

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Claas T. Buschmann 1 ,
Biagio Solarino 2 &
Takahito Hayashi 3

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After nearly 15 years, the scientific Chief Editorial Team of Forensic Science, Medicine and Pathology has changed in January 2022 [ 1 , 2 ].

First of all, we gratefully thank former Editor-in-Chief Roger W. Byard, former European Editor Michael Tsokos, and former North American Editor John Hunsaker III for their time and passion invested over the past 15 years in the journal — they have not only read, reviewed, and edited thousands of papers, they have also published extensively themselves and thus contributed to the reputation of the journal, too. Together with the Editorial Board, they made the journal what it is today.

Secondly, we would like to thank the whole team at Springer, who gave us three “newcomers” the opportunity to follow in these big footsteps. We will do our best.

Now brought to the second-most important journal worldwide in forensic medicine with regard to its current impact factor by the former Editorial team, Forensic Science, Medicine and Pathology will continue to explore all aspects of modern-day forensics. The range of topics covered will continue to include international forensic science, medicine, nursing, and pathology, as well as toxicology, human identification, mass disasters/mass war graves, profiling, imaging and forensic radiology, forensic age estimation, policing, wound assessment, child maltreatment, sexual assault, anthropology, archeology, entomology, botany, biology, veterinary pathology, medical-historical forensic research, and DNA. We will continue to insist on high scientific quality of papers in fluent and sufficiently readable English language. We pay great attention to evaluating the mega-authorship reports and ask to limit the number of self-citations, particularly if not necessary.

Mors auxilium vitae (Death Is Help For The Living) , and looking beyond the horizon is crucial in modern forensic medicine. Interdisciplinary questions arising in the daily autopsy routine can be addressed scientifically, and forensic medicine can contribute to walk new paths. Thus, we also welcome “outside-the-box” papers, i.e., scientific research from the interface of forensic medicine and other medical disciplines — there is a significant overlap between forensic medicine and several curative disciplines, especially after a second look [ 3 , 4 , 5 ]. This also applies to the interface of forensic medicine and the judicial system, i.e., legal assessment of forensic findings. As a sub-category of case reports, we have established “From The Court Room” as a brief case description to present and discuss — not necessarily extraordinary — autopsy and/or crime scene features in a specific case and their legal evaluation. What are legal consequences of our work for those affected, and where are the limits of forensic diagnostics? Where can we get better? With this new proposal, we can discuss once again the inference of the forensic publications in a trial [ 6 ]. How does the Judge determine the scientific value of the articles and the qualifications and credentials of a proposed expert witness? Even considering the differences among the legal systems worldwide, we are looking forward to submissions addressing these points.

Forensic Science, Medicine and Pathology will continue to present a balance of forensic research and reviews from around the world to reflect modern advances through peer-reviewed papers, short communications, meeting proceedings, new forensic textbook comments, and case reports. Furthermore, we will open the journal to answers to forensic questions that involve interfaces with other medical disciplines, especially with regard to complications arising from performed — or necessary, but omitted — medical procedures in the broadest sense. Forensic scientists are often involved in medical malpractice lawsuits, healthcare policy, and patient safety management. Therefore, the authors have the opportunity to discuss unusual adverse events, causes of medical malpractice, and the forensic medicine approach to such an interesting field of research. The dead can teach the living.

The scientific future of forensic medicine comprises not only of forensic issues, but involves interdisciplinary cooperation. We intend to be a relevant part of this future — and we can achieve this goal solely with the help of you, the authors and reviewers from all over the world!

Priv.-Doz. Dr. med. Claas T. Buschmann, Kiel/Lübeck, Germany

Editor-in-Chief

Prof. Dr. Biagio Solarino, Bari, Italy

Associate Editor

Prof. Takahito Hayashi, Kagoshima, Japan

Byard RW, Hunsaker JC, Tsokos M. Forensic science medicine and pathology – a change of command. Forensic Sci Med Pathol. 2022;18:116–7.

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Byard RW. Academic standing and publication. Forensic Sci Med Pathol. 2022;18:1–3.

Buschmann C. More interdisciplinary research is needed in forensic medicine. Forensic Sci Med Pathol. 2019;15:131–2.

Buschmann C, Tsokos M, Kleber C. Preventive pathology: the interface of forensic medicine and trauma surgery for pre-hospital trauma management. Forensic Sci Med Pathol. 2015;11:317–8.

Lacour P, Buschmann C, Storm C, et al. Cardiac implantable electronic device interrogation at forensic autopsy – an underestimated resource? Circulation 2018;137:2730–40.

Jones AW. Highly cited forensic practitioners in the discipline legal and forensic medicine and the importance of peer-review and publication for admission of expert testimony. Forensic Sci Med Pathol. 2022;18:37–44.

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Institute of Legal Medicine, University Hospital Schleswig-Holstein, Kiel / Lübeck, Germany

Claas T. Buschmann

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Biagio Solarino

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Buschmann, C.T., Solarino, B. & Hayashi, T. Big footsteps and new challenges. Forensic Sci Med Pathol 18 , 123–124 (2022). https://doi.org/10.1007/s12024-022-00482-5

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NIJ Announces New Forensic Science Strategic Research Plan for 2022–2026

NIJ Forensic Science Strategic Research Plan 2022-2026

June 22, 2022

The National Institute of Justice (NIJ) Office of Investigative and Forensic Sciences has published its latest four-year Forensic Science Strategic Research Plan .

The plan focuses on improving the quality and practice of forensic science through innovative solutions that support research and development, testing and evaluation, technology, and information exchange.

As stated in the plan, “NIJ developed this Forensic Science Strategic Research Plan to communicate its research agenda and advance its forensic science research mission. The strategic priorities and objectives outlined in this plan closely parallel the opportunities and challenges faced by the forensic science community.”

NIJ identified five strategic research priorities:

Strategic Priority 1: Advance Applied Research and Development in Forensic Science
Strategic Priority 2: Support Foundational Research in Forensic Science
Strategic Priority 3: Maximize the Impact of Forensic Science Research and Development
Strategic Priority 4: Cultivate a Diverse, Highly Skilled Forensic Science Workforce
Strategic Priority 5: Coordinate Across the Community of Practice

Each priority includes a comprehensive set of objectives and action items.

According to the NIJ, the strategic research plan should interest many stakeholders, including crime laboratory professionals, researchers and technology developers, the legal community, and policymakers.

The Forensic Science Strategic Research plan is available to download at https://nij.ojp.gov/topics/articles/forensic-sciences-strategic-research-plan-2022-2026 .

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Recent advances in forensic biology and forensic DNA typing: INTERPOL review 2019–2022

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This review paper covers the forensic-relevant literature in biological sciences from 2019 to 2022 as a part of the 20th INTERPOL International Forensic Science Managers Symposium. Topics reviewed include rapid DNA testing, using law enforcement DNA databases plus investigative genetic genealogy DNA databases along with privacy/ethical issues, forensic biology and body fluid identification, DNA extraction and typing methods, mixture interpretation involving probabilistic genotyping software (PGS), DNA transfer and activity-level evaluations, next-generation sequencing (NGS), DNA phenotyping, lineage markers (Y-chromosome, mitochondrial DNA, X-chromosome), new markers and approaches (microhaplotypes, proteomics, and microbial DNA), kinship analysis and human identification with disaster victim identification (DVI), and non-human DNA testing including wildlife forensics. Available books and review articles are summarized as well as 70 guidance documents to assist in quality control that were published in the past three years by various groups within the United States and around the world.

1. Introduction

This review explores developments in forensic biology and forensic DNA analysis of biological evidence during the years 2019–2022. In some cases, there may be overlap with 2019 articles mentioned in the previous INTERPOL review covering 2016 to 2019 [ 1 ]. This review includes books and review articles, published guidance documents to assist in quality control, rapid DNA testing, using law enforcement DNA databases plus investigative genetic genealogy DNA databases along with privacy/ethical issues, forensic biology and body fluid identification, DNA extraction and typing methods, mixture interpretation involving probabilistic genotyping software (PGS), DNA transfer and activity level evaluations, next-generation sequencing (NGS), DNA phenotyping, lineage markers (Y-chromosome, mitochondrial DNA, X-chromosome), new markers and approaches (microhaplotypes, proteomics, and microbial DNA), kinship analysis and human identification with disaster victim identification (DVI), and non-human DNA testing including wildlife forensics.

Multiple searches, using the Scopus (Elsevier) and Web of Science (Clarivate) databases, were conducted in the first half of 2022 with “forensic” and “DNA” or “biology” and “2019 to 2022” as search options. Over 4000 articles were returned with these searches. Through visual examination of titles and authors, duplicates were removed, and articles sorted into 32 subcategories to arrive at a list of almost 2000 publications that were supplemented throughout the remainder of the year as this review was being prepared. The tables of contents for non-indexed journals, such as WIRES Forensic Science , Journal of Forensic Identification , and Forensic Genomics were also examined to locate potentially relevant articles.

For example, a Scopus search conducted on June 13, 2022, using “forensic DNA” and “2019 to 2022” found a total of 3059 documents. Table 1 lists the top ten journals from this search. The Forensic Science International: Genetics Supplement Series (see row #4 in Table 1 ) provides the proceedings of the International Society for Forensic Genetics (ISFG) meeting held in Prague in September 2019. This volume contains 914 pages with 347 articles (although only 172 showed up in the Scopus search) that are freely available at https://www.fsigeneticssup.com /[ 2 ]. Thus, searches conducted with one or even multiple databases (e.g., Scopus and Web of Science) may not be comprehensive or exhaustive.

Top ten journals with forensic DNA articles published from 2019 to 2022 based on a Scopus search on June 13, 2022.

1.1. Books, special issues, and review articles of note

Books published during the period of this review relating to forensic biology and forensic DNA include Essential Forensic Biology, Third Edition [ 3 ], Principles and Practices of DNA Analysis: A Laboratory Manual for Forensic DNA Typing [ 4 ], Forensic DNA Profiling: A Practical Guide to Assigning Likelihood Ratios [ 5 ], Forensic Practitioner's Guide to the Interpretation of Complex DNA Profiles [ 6 ], Silent Witness: Forensic DNA Evidence in Criminal Investigations and Humanitarian Disasters [ 7 ], Mass Identifications: Statistical Methods in Forensic Genetics [ 8 ], Probability and Forensic Evidence: Theory, Philosophy, and Applications [ 9 ], Interpreting Complex Forensic DNA Evidence [ 10 ], Understanding DNA Ancestry [ 11 ], Understanding Forensic DNA [ 12 ], and Handbook of DNA Profiling [ 13 ]. The 2022 Handbook of DNA Profiling spans two volumes and 1206 pages with 54 chapters from 115 contributors representing 17 countries.

Over the past three years, several special issues on topics related to forensic biology were published in Forensic Science International: Genetics and Genes . These special issues were typically collated virtually rather than physically as invited articles were published online over some period of time and then bundled together virtually as a special issue. Some of these review articles or a set of special issue articles are open access (i.e., the authors paid a publication fee so that the article would be available online for free to readers).

During the time frame of this INTERPOL DNA review, FSI Genetics published two special issues: (1) “Trends and Perspectives in Forensic Genetics” (editor: Manfred Kayser) 1 with nine review and two original research articles published between September 2018 and January 2019, and (2) “Forensic Genetics – Unde venisti et quo vadis?” [Latin for “where did you come from and where are you going?”] (editor: Manfred Kayser) with nine articles published in 2021 and early 2022 and likely two more before the end of 2022. Topics for review articles in these special issues include DNA transfer [ 14 ], probabilistic genotyping software [ 15 ], microhaplotypes in forensic genetics [ 16 ], investigative genetic genealogy [ 17 ], forensic proteomics [ 18 ], distinguishing male monozygotic twins [ 19 ], and using the human microbiome for estimating post-mortem intervals and identifying individuals, tissues, or body fluids [ 20 , 21 ]. All of these topics will be discussed later in this article.

A Genes special issue “Forensic Genetics and Genomics” (editors: Emiliano Giardina and Michele Ragazzo) 2 published 11 online articles plus an editorial from April 2020 to January 2021 while another Genes special issue “Forensic Mitochondrial Genomics” (editors: Mitch Holland and Charla Marshall) 3 compiled 11 articles from February 2020 to April 2021. An “Advances in Forensic Genetics” Genes special issue (editor: Niels Morling) 4 included 25 articles shared between April 2021 and May 2022. In July 2022, the Advances in Forensic Genetics articles were compiled as a 518-page book. 5 Other Genes special issues in development or forthcoming covering aspects of forensic DNA and requesting potential manuscripts by late 2022 or early 2023 include “State-of-the-Art in Forensic Genetics” (editor: Chiara Turchi), 6 “Trends in Population Genetics and Identification—Impact on Anthropology (editors: Antonio Amorim, Veronica Gomes, Luisa Azevedo), 7 “Identification of Human Remains for Forensic and Humanitarian Purposes: From Molecular to Physical Methods” (editors: Elena Pilli, Cristina Cattaneo), 8 “Improved Methods in Forensic and DNA Analysis” (editor: Marie Allen), 9 “Forensic DNA Mixture Interpretation and Probabilistic Genotyping” (editor: Michael Coble) 10 , and “Advances in Forensic Molecular Genetics” (editors: Erin Hanson and Claire Glynn). 11 There has been a proliferation of review articles and special issues in this field in the past several years!

A new journal Forensic Science International: Reports was launched in November 2019. As of June 2022, it has published 89 articles involving DNA, most of which are descriptions of population genetic data. Likewise, a June 27, 2022, PubMed search with “forensic DNA” and the journal “Genes” found 88 articles – many of which are part of the previously mentioned special issues.

1.2. Guidance documents

Numerous documentary standards and guidance documents related to forensic DNA have been published by various organizations around the world. Table 2 lists 70 such documents released in the past three years (2019–2022) in the United States, UK, Australia, and the European Union.

Guidance documents related to forensic DNA published from 2019 to 2022. The titles are hyperlinked to available documents. Abbreviations: FBI (Federal Bureau of Investigation), CODIS (Combined DNA Index System), SWGDAM (Scientific Working Group on DNA Analysis Methods), NGS (next generation sequencing), US DOJ (United States Department of Justice), ULTR (Uniform Language for Testimony and Reports), AABB (Association for the Advancement of Blood and Biotherapies), ASB (Academy Standards Board), OSAC (Organization of Scientific Area Committees for Forensic Science), UKFSR (United Kingdom Forensic Science Regulator), ENFSI (European Network of Forensic Science Institutes), NIFS (National Institute of Forensic Science), ISFG (International Society for Forensic Genetics).

1.2.1. SWGDAM, FBI, and other US DOJ activities

The Federal Bureau of Investigation (FBI) Laboratory funds the Scientific Working Group on DNA Analysis Methods (SWGDAM) 12 to serve as a forum for discussing, sharing, and evaluating forensic biology methods, protocols, training, and research. In addition to creating guidelines on various topics, SWGDAM, which meets semiannually in January and July, provides recommendations to the FBI Director on the Quality Assurance Standards (QAS) used to assess U.S. forensic DNA laboratories involved in the National DNA Index System (NDIS) that perform DNA databasing and forensic casework. New versions of the QAS became effective July 1, 2020.

SWGDAM work products from the timeframe of 2019–2022 (see Table 2 ) include QAS audit and guidance documents, mitochondrial DNA analysis and short tandem repeat (STR) interpretation guideline revisions related to next-generation sequencing (NGS), training and Y-chromosome interpretation guidelines, a Y-chromosome Haplotype Reference Database (YHRD) update for U.S. laboratories, and reports on investigative genetic genealogy and Y-screening of sexual assault evidence kits. These documents are all accessible online. 13

In January 2022, the FBI produced a 13-page guide 14 on rapid DNA testing describing booking station applications and their vision for future integration of crime scene sample analysis and the Combined DNA Index System (CODIS), which builds on a joint position statement published in July 2020 by leaders of U.S. and European groups [ 22 ]. In addition, the FBI has shared guidance on their website for non-CODIS use of rapid DNA testing with law enforcement applications 15 and considerations for court. 16

United States Department of Justice (US DOJ) Uniform Language for Testimony and Reports (ULTRs), 17 contain three ULTRs for the forensic DNA discipline that became effective in March 2019: autosomal DNA with probabilistic genotyping, mitochondrial DNA, and Y-STR DNA. USDOJ also released an interim policy on investigative genetic genealogy in November 2019 [ 23 ] along with an opinion piece in the journal Science calling for responsible genetic genealogy [ 24 ].

Other agencies within US DOJ, namely the Bureau of Justice Assistance (BJA) and the National Institute of Justice (NIJ), published a guide for prosecutors on triaging forensic evidence [ 25 ] and best practices for improving DNA laboratory process efficiency [ 26 ]. A 200-page report to Congress on the needs assessment of forensic laboratories and medical examiner/coroner offices was released in December 2019 calling for $640 million annually in additional funding to support U.S. forensic efforts [ 27 ].

In September 2021, the Forensic Technology Center of Excellence (FTCOE), which is funded by NIJ, published a 29-page implementation strategy on next-generation sequencing for DNA analysis that was written by the NIJ Forensic Laboratory Needs Technology Working Group (FLN-TWG) [ 28 ]. In May 2022, FTCOE released a 50-page landscape study examining technologies and automation for differential extraction and sperm separation used in sexual assault investigations [ 29 ]. An introduction to forensic genetic genealogy was released in September 2022 [ 30 ].

The FTCOE also published a human factors forensic science sourcebook 18 in March 2022 through open access articles in the journal Forensic Science International: Synergy . This sourcebook, which has general applicability rather than being specific to forensic DNA analysts, includes an overview article [ 31 ] along with articles on personnel selection and assessment [ 32 ], the benefits of committing errors during training [ 33 ], how characteristics of human reasoning and certain situations can contribute to errors [ 34 ], stressors that impact performance [ 35 ], and the impact of communication between forensic analysts and detectives using a new metaphor [ 36 ].

1.2.2. OSAC and ASB activities

The Organization of Scientific Area Committees for Forensic Science (OSAC) 19 is congressionally-funded and administered by the Special Programs Office within the National Institute of Standards and Technology (NIST). OSAC consists of a governing board and over 600 members and associates organized into seven scientific area committees (SACs) and 22 subcommittees. The Biology SAC is divided into human and wildlife forensic biology activities. The Human Forensic Biology Subcommittee 20 focuses on standards and guidelines related to training, method development and validation, data analysis, interpretation, and statistical analysis as well as reporting and testimony for human forensic serological and DNA testing. The Wildlife Forensics Subcommittee 21 works on standards and guidelines related to taxonomic identification, individualization, and geographic origin of non-human biological evidence based on morphological and genetic analyses.

The Academy Standards Board (ASB) 22 is a wholly owned subsidiary of the American Academy of Forensic Sciences (AAFS) and was established as a standards developing organization (SDO). In 2015, ASB was accredited as an SDO by the American National Standards Institute (ANSI). The ASB DNA Consensus Body, with a membership consisting of practitioners, researchers, and lawyers, develops standards and guidelines related to the use of DNA in legal proceedings. Many of the documents developed by ASB were originally proposed OSAC standards or guidelines.

The OSAC Registry 23 is a repository of high-quality and technically-sound standards (both published and proposed) that are intended for implementation in forensic science laboratories. As of July 2022, the OSAC Registry contains 11 standards published by ASB as well as two (2) proposed OSAC standards or best practice recommendations related to human forensic biology. Another four ASB standards and two proposed OSAC standards related to wildlife forensic biology are on the OSAC Registry. The ASB standards issued in the past three years related to human forensic biology cover interpretation and comparison protocols, training in various parts of the process, and validation of forensic serological and DNA analysis methods as well as probabilistic genotyping systems (see Table 2 for names of these documents). A number of other documents 24 related to serological testing methods, assigning propositions for likelihood ratios in forensic DNA interpretations, validation of forensic DNA methods and software, familial DNA searching, management and use of quality assurance DNA elimination databases, setting thresholds, evaluative forensic DNA testimony, and training in use of statistics are in development within OSAC and ASB.

Additional work products of OSAC include (1) a lexicon 25 with 3282 records (although multiple records may exist for the same word, e.g., there are five definitions provided for “validation” from various sources), (2) a 35-page technical guidance document 26 on human factors in validation and performance testing that describes key issues in designing, conducting, and reporting validation research, (3) a listing of research and development needs in forensic science 27 including 18 identified by the OSAC Human Forensic Biology Subcommittee during their deliberations ( Table 3 ), and (4) process maps for several forensic disciplines including a 42-page depiction of current practices and decisions in human forensic DNA analysis released in May 2022 [ 37 ]. As a visual representation of critical steps and decision points, a process map is intended to help improve efficiencies and reduce errors, and highlight gaps where further research or standardization would be beneficial. Process maps can assist with training new examiners and enable development of specific laboratory policies or help identify best practices for the field.

Research and development needs in forensic biology as identified by the OSAC Human Forensic Biology Subcommittee (as of July 2022, see https://www.nist.gov/osac/osac-research-and-development-needs ).

1.2.3. UK Forensic Science Regulator

The UK Forensic Science Regulator (UKFSR) oversees forensic science efforts in England, Wales, and Northern Ireland. In March 2021, the Regulator released the seventh issue 28 of the Codes of Practice and Conduct for forensic science providers and practitioners in the criminal justice system. This 114-page document, which has been updated every few years, provides the overall framework for forensic science activities in the UK with other supporting guidance documents on specific areas like DNA analysis or general tasks like validation. In September 2020, a number of the Regulator documents were revised and reissued. As noted in Table 2 (see rows with documents containing “Issue 1” in the title), new guidance documents were also released in the past few years on sexual assault examinations, development of evaluative opinions, proficiency testing for DNA mixture interpretation, Y-STR profiling, DNA relationship testing, and methods employing rapid DNA testing devices. Table 2 lists 20 guidance documents pertinent to forensic biology from the UKFSR.

1.2.4. European Union and Australia

The European Network of Forensic Science Institutes (ENFSI) DNA Working Group published two documents in the past three years: one on DNA database management and the other on training of staff in forensic DNA laboratories (see Table 2 ). A best practice manual for human forensic biology and DNA profiling is also under development.

The Australian National Institute of Forensic Science (NIFS) published three documents of relevance to forensic biology on case record review, empirical study design, and transitioning technology from the laboratory to the field (see Table 2 ).

1.2.5. Other international efforts

The Association for the Advancement of Blood and Biotherapies (AABB) 29 published the 15th edition of their Standard for Relationship Testing Laboratories, which became effective on January 1, 2022. This documentary standard was developed by the AABB Relationship Testing Standards Committee and applies to laboratories accredited for paternity testing and other forms of genetic relationship assessment.

The International Society for Forensic Genetics (ISFG) DNA Commission 30 published two articles during the timeframe of this INTERPOL review (see Table 2 ). In 2020, guidelines and considerations were published on evaluating DNA results under activity level propositions [ 38 ]. In addition, the state of the field regarding interpretation of Y-STR results was examined along with different approaches for haplotype frequency estimation using population data – with the Discrete Laplace approach being recommended [ 39 ]. Future ISFG DNA Commission efforts will address STR allele sequence nomenclature and phenotyping.

2. Advancements in current practices

This section (Section 2 ) is intended to be law enforcement and practitioner-focused through examination of advances in current practices. The following section (Section 3 ) is intended to be researcher-focused through emphasis on emerging technologies and new developments. In this section, topics specifically covered include rapid DNA analysis, use of DNA databases to aid investigations (including familial searching, investigative genetic genealogy, genetic privacy and ethical concerns, and sexual assault kit testing), body fluid identification, DNA extraction and typing methods, and DNA interpretation at the sub-source and activity level.

2.1. Rapid DNA analysis

Rapid DNA instruments that provide integrated “swab-in-profile-out” results in 90 min or less can be used in police booking station environments and assist investigations outside of a traditional laboratory environment. These instruments were initially designed for analysis of buccal swabs to help speed processing of reference samples associated with criminal cases. Such samples are expected to contain relatively large quantities of DNA from a single contributor. Some attempts to extend the range of sample types to low quantities of DNA or mixtures have been published with various levels of success (see Table 4 ). Researcher and practitioners from Australia [ [40] , [41] , [42] ], Canada [ 43 ], China [ 44 ], Italy [ 45 ], Japan [ 46 , 47 ], and the United States [ [48] , [49] , [50] , [51] , [52] , [53] , [54] , [55] , [56] , [57] ] have contributed to an increased understanding of rapid DNA testing capabilities and limitations.

Summary of 20 rapid DNA instrument validation and evaluation studies published from 2019 to 2022. Abbreviations: A-Chip (arrestee cartridge, designed for high-quantity DNA samples), I-Chip (investigative cartridge, designed for low-quantity DNA samples), ACE (arrestee cartridge with GlobalFiler STR markers), RapidINTEL (uses 32 rather than 28 PCR cycles to increase success with low-quantity DNA samples). A-Chip and I-Chip amplify the FlexPlex set of 23 autosomal STRs, three Y-STRs, and amelogenin [ 51 ]. ACE and RapidINTEL utilize the GlobalFiler set of 21 autosomal STRs, one Y-STR, one Y-chromosome InDel, and amelogenin.

The Accelerated Nuclear DNA Equipment (ANDE) 6C (ANDE, Longmont, CO, USA) and the RapidHIT ID (Thermo Fisher Scientific, Waltham, MA, USA) are the current 31 commercially available rapid DNA systems. Each system consists of a swab for introducing the sample, a cartridge or biochip with pre-packed reagents, the instrument, and analysis software with an expert system for automated STR allele calling. Different sample cartridges can be run on each system depending on the sample type and expected quantity of DNA.

For ANDE, the arrestee cartridge (A-Chip), can accommodate up to five samples and is intended for relatively high quantities of DNA typically collected from reference buccal swabs, while the investigative cartridge (I-Chip), can process up to four samples and is intended for lower quantities of DNA that might be present in casework or disaster victim identification samples. Both ANDE cartridges use the FlexPlex27 STR assay that tests 23 autosomal STR loci, three Y-chromosome STRs, and amelogenin to generate data compatible with DNA databases around the world [ 51 ]. The RapidHIT ID ACE cartridge and RapidINTEL cartridge serve similar purposes as the ANDE A-Chip and I-Chip using GlobalFiler Express kit markers (21 autosomal STRs, DYS391, a Y-chromosome insertion/deletion marker, and amelogenin) instead of the FlexPlex assay. The ACE sample cartridge uses buccal swabs while the EXT sample cartridge processes DNA extracts [ 56 ]. Sensitivity is enhanced in the RapidINTEL cartridge by increasing the number of PCR cycles from 28 to 32 and decreasing the lysis buffer volume from 500 μL to 300 μL compared to the ACE cartridge parameters [ 46 ].

With rapid DNA testing's swab-in and answer-out integrated configuration, limited options exist for testing conditions (e.g., either A-Chip or I-Chip with ANDE). Therefore, users should evaluate performance for the sample types they desired to routinely test in their specific environment. Table 4 summarizes recently published studies containing rapid DNA assessments.

National DNA Index System (NDIS) approval has been provided by the FBI Laboratory for accredited forensic DNA laboratories to use either the ANDE 6C or RapidHIT ID Systems (A-Chip and ACE cartridges only) 32 with eligible reference mouth swabs. As noted in Table 2 , the FBI.gov website contains three documents related to rapid DNA testing: “Non-CODIS Rapid DNA Considerations and Best Practices for Law Enforcement Use” (7-pages), “Rapid DNA Testing for Non-CODIS Uses: Considerations for Court” (5-pages), and “A Guide to All Things Rapid DNA” (13-pages) in January 2022 to provide information on the topic to law enforcement agencies.

The ENFSI DNA Working Group, SWGDAM, and an FBI Rapid DNA Crime Scene Technology Advancement Task Group co-published a position statement on the use of rapid DNA testing from crime scene samples [ 22 ]. These groups emphasized the need to have future rapid DNA systems with (1) methods to identify low quantity, degradation, and inhibition as well as meeting the human quantification requirements shared by SWGDAM and others, (2) the ability to export analyzable raw data for analysis or reanalysis by trained and qualified forensic DNA analysts, (3) an on-board fully automated expert system to accurately flag single-source or mixture DNA profiles requiring analyst evaluation, (4) improved peak height ratio balance (per locus and across loci) for low-quality and mixture samples “through enhancements in extraction efficiencies, changes in cycling parameters, and/or changes in STR kit chemistries,” and (5) published developmental validation studies on a wide variety of forensic evidence type samples with “data-supported recommendations regarding types of forensic evidence that are suitable and unsuitable for use with Rapid DNA technology” [ 22 ].

With a likely increase in the capabilities and the availability of rapid DNA systems, investigators will need to decide whether to use this capability onsite in specific situations or to send collected samples to a conventional forensic laboratory for processing at a later time. A group in the Netherlands collaborated with the New York City Police Department Crime Scene Unit and Evidence Collection Team to explore a decision support system [ 60 ]. In this study, participants were informed that rapid DNA testing was less sensitive compared to laboratory analysis and that the sample would be consumed, but that results from rapid DNA testing could identify a suspect within 2 h as opposed to waiting an average of 45 days for the laboratory results [presumably due to sample backlogs]. They were also told that a DNA profile obtained with rapid DNA would be acceptable in court. In the end, “>90% of the participants (85 out of 91) saw added value for using a Rapid DNA device in their investigative process …” with “a systematic approach, which consists of weighing all possible outcomes before deciding to use a Rapid DNA analysis device” [ 60 ]. The authors note that for such an approach to be successful “knowledge on DNA success rates [with various evidence types] is necessary in making evidence-based decisions for Rapid DNA analysis” [ 60 ].

A group in Australia performed a cost-benefit analysis of a decentralized rapid DNA workflow that might exist in the future with instruments placed at police stations around their country [ 61 ]. A virtual assessment considered all reference DNA samples collected during a two-month time period at 10 participating police stations in five regions of Australia. Processing times at the corresponding DNA analysis laboratories were calculated based on when the sample was received compared to the day when a DNA profile was obtained for that sample. From the survey conducted, it was estimated that up to 80,000 reference DNA samples are currently processed each year in forensic DNA laboratories across Australia [ 61 ].

Consumable costs for conventional DNA testing reagents in Australia were found to range from $17 to $35 whereas the rapid DNA consumable costs were estimated to be $100 per sample along with an anticipated $100,000 instrument cost per police station. Of course, the rate of use is expected to vary based on the number of reference samples collected in that jurisdiction. Since rapid DNA instruments utilize consumable cartridges with expiration dates, it was estimated that a police station would need to process six DNA samples per week to avoid having to discard an expired cartridge and thus increase the overall cost of their rapid DNA testing efforts. The authors of this study conclude “that routine laboratory DNA analysis meets the current needs for the majority of cases … It is anticipated that while the cost discrepancy between laboratory and rapid DNA processing remains high, the uptake of the technology in Australia will be limited [at least for a police booking station scenario]” [ 61 ].

Rapid DNA technology can be used in a variety of contexts including some that extend beyond traditional law enforcement. Seven distinct use contexts for rapid DNA capabilities have been described [ 62 ]: (1) evidence processing at or near crime scenes to generate leads for confirmation by a forensic laboratory, (2) booking or detection stations to compare an individual's DNA profile to a forensic database while the individual is still in custody, (3) disaster victim identification to permit rapid DNA processing of a victim's family members during their visit to family assistance centers when filing missing persons reports, (4) missing persons investigations to quickly process unidentified human remains and/or family reference samples to generate leads for confirmation by a forensic laboratory, (5) border security to develop DNA data from detainees for comparison to indices of prior border crossers while the individual is still in custody, (6) human trafficking and immigration fraud detection to permit immigration officials to verify family relationship claims, and (7) migrant family reunification to allow immigration officials to verify parentage claims and reunite family members separated at the border. Social and ethical considerations have been proposed for each of these use contexts in terms of data collection, data access and storage, and oversight and data protection [ 62 ].

One study [ 47 ] evaluating buccal swabs and mock disaster victim identification samples drew an important conclusion worth repeating here: “The Rapid DNA system provides robust and automated analysis of forensic samples without human review. Sample analysis failure can happen by chance in both the Rapid DNA system and conventional laboratory STR testing. While re-injection of PCR product is easily possible in the conventional method, this is not an option with the Rapid DNA system. Accordingly, the Rapid DNA system is a suitable choice but should be limited to samples that can easily be collected again if necessary or to samples that are of sufficient amount for repeated analysis. Application of this system to valuable samples such as those related to casework need to be considered carefully before analysis.”

2.2. Using DNA databases to aid investigations (national databases, familial searching, investigative genetic genealogy, genetic privacy & ethical concerns, sexual assault kit testing)

Forensic DNA databases can aid investigations by demonstrating connections between crime scenes, linking a previously enrolled DNA profile from an arrestee or convicted offender to biological material recovered from a crime scene, or aiding identification of missing persons through association of remains with biological relatives. Establishment of these databases requires significant investments over time to enroll data from crime scenes and potential serial offenders or unidentified human remains and relatives of missing persons. This section explores issues around national DNA databases, familial searching, investigative genetic genealogy, and genetic privacy and ethical concerns.

A systematic review regarding the effectiveness of forensic DNA databases looked at 19 articles published between 1985 and 2018 and found most studies support the assumption that DNA databases are an effective tool for the police, society, and forensic scientists [ 63 ]. Recommendations have been proposed to make cross-border exchange of DNA data more transparent and accountable with the Prüm system that enables information sharing across the European Union [ 64 ]. An analysis of news articles discussing the use of DNA testing in family reunification with migrants separated at the U.S.-Mexico border has been performed [ 65 ], and a standalone humanitarian DNA identification database has been proposed [ 66 ]. Aspects of international DNA kinship matching were explored to aid missing persons investigations and disaster victim identification processes [ 67 ]. A business case was presented for expanded DNA indirect matching using additional genetic markers, such as Y-chromosome STRs, mitochondrial DNA, and X-chromosome STRs, to reveal previously undetected familial relationships [ 68 ].

Approaches to transnational exchange of DNA data include (1) creation of an international DNA database, (2) linked or networked national DNA databases, (3) request-based exchange of data, and (4) a combination of these [ 69 ]. For example, the INTERPOL DNA database 33 contains more than 247,000 profiles contributed by 84 member countries. The I-Familia global database assists with missing persons identification based on international DNA kinship matching. 34

2.2.1. National DNA databases

Since the United Kingdom launched the first national DNA database in 1995, national DNA databases continue to be added in many countries including Brazil [ 70 , 71 ], India [ 72 ], Pakistan [ 73 , 74 ], Portugal [ 75 ], and Serbia [ 76 ]. A survey of 15 Latin American countries found that 13 of them had some kind of DNA database [ 77 ]. The opinions of 210 prisoners and prison officials in three Spanish penitentiary centers were also collected regarding DNA databases [ 78 ].

The effectiveness of databases has been debated over the years. Seven key indicators were used in a 2019 examination of the effectiveness of the UK national DNA database. These indicators included (1) implementation cost – the financial input required to implement the database system, (2) crime-solving capability – the ability of the database to assist criminal justice officials in case resolution, (3) incapacitation effect – the ability of the database to reduce crime through the incapacitation of offenders, (4) deterrence effect – the preventative potential of the database through deterrence of individuals from committing crime, (5) privacy protection – protection of the privacy or civil liberty rights of individuals, (6) legitimacy – compliance of the databasing system to the principle of proportionality, and (7) implementation efficiency – the time and non-monetary resource required to implement the database system [ 79 ].

A follow-up article concluded: “Available evidence shows that while DNA analysis has contributed to successful investigations in many individual cases, its aggregate value to the resolution of all crime is low” [ 80 ]. The systematic review of 19 articles on DNA databases cited previously noted “the expansion of DNA databases would only have positive effects on detection and clearance if the offender were already included in the database” [ 63 ]. When previous offenders are not already in a law enforcement DNA database to provide a hit to a crime scene profile, efforts are increasingly turning to familial searching and investigative genetic genealogy as described in the following sections.

2.2.2. Familial DNA searching

Familial DNA searching (FDS) extends the traditional direct matching of STR profiles within law enforcement databases to search for potential close family relationships, such as a parent or sibling, of a profile in the database. 35 FDS typically uses Y-STR lineage testing to narrow the set of candidate possibilities along with other case information such as geographic details of the crime and age of the person(s) of interest. For example, FDS helped solve murder cases in Romania [ 81 ] and China [ 82 ] by locating the perpetrator through a relative in the DNA database. A survey of 103 crime laboratories in the United States found that 11 states use FDS while laboratories in 24 states use a similar but distinct practice of partial matching [ 83 ].

The expansion of the number of STRs from 15 to 20 or 21 helps distinguish between true and false matches during a DNA database search by reducing the number of FDS adventitious matches [ 84 ]. Another study noted that the choice of allele frequencies affects the rate at which non-relatives are erroneously classified as relatives and found that using ancestry inference on the query profile can reduce false positive rates [ 85 ]. New Y-STR kits have been developed to assist with familial searching [ 86 , 87 ]. FDS of law enforcement databases differs from investigative genetic genealogy in two important ways – the genetic markers and the databases used for searching [ 88 , 89 ].

2.2.3. Investigative genetic genealogy

In recent years when national DNA databases fail to generate a lead to a potential person of interest, law enforcement agencies have started to utilize the capabilities of investigative genetic genealogy (IGG), also called forensic genetic genealogy (FGG) or forensic investigative genetic genealogy (FIGG), as an approach to locate potential persons of interest in criminal or missing persons cases. For example, a pilot case study in Sweden used IGG to locate the perpetrator of a double murder from 2004 who had evaded detection despite 15 years of various investigation efforts including more than 9000 interrogations and mass DNA screenings of more than 6000 men [ 90 ]. Hardly a week goes by without mention in the global media of another cold case being solved with IGG. Since the arrest of Joseph DeAngelo in April 2018 identified as the infamous Golden State Killer using IGG, hundreds of cold criminal and unidentified human remains cases have been resolved [ 91 ].

IGG involves examination of about 600,000 single nucleotide polymorphisms (SNPs), rather than the 20 or so STRs used in conventional forensic DNA testing, to enable associations of relatives as distant as third or fourth cousins [ 17 ]. IGG relies on a combination of publicly accessible records and the consent of individuals who have uploaded their genetic genealogy DNA profiles to genetic genealogy databases [ 92 ]. Multiple reviews and research articles have been published describing current IGG methods, knowledge, and practice along with the effectiveness and operational limits of the technique [ 17 , 30 , [93] , [94] , [95] , [96] , [97] ]. IGG works best with high-quality, single-source DNA samples. A case study involving whole genome sequencing of human remains from a 2003 murder victim found that it was possible to perform IGG for identification of the victim in this situation [ 98 ].

The four main direct-to-consumer (DTC) genetic genealogy companies, 23andMe (Mountain View, CA), Ancestry (Salt Lake City, UT), FamilyTree DNA (Houston, TX), and My Heritage (Lehi, UT), have DNA data from over 41 million individuals 36 as of July 2022 [ 97 ]. Individuals can upload their DTC data to GEDmatch, which is a DNA comparison and analysis website launched in 2010 and purchased in 2019 by Verogen (San Diego, CA). Law enforcement IGG searches are currently permitted with DTC data for individuals who opt into the GEDmatch database or do not opt out of the FamilyTree DNA database [ 99 , 100 ]. Currently most DTC genetic genealogy data comes from the United States and individuals of European origin. A UK study found that 4 of 10 volunteer donors could be identified with IGG including someone of Indian heritage demonstrating that under the right circumstances individuals of non-European origin can be identified [ 101 ].

As noted previously in Section 1.2.1 , the U.S. Department of Justice released an interim policy guide to forensic genetic genealogical DNA analysis and searching [ 23 ], and the FBI Laboratory's chief biometric scientist published an editorial in Science calling for responsible genetic genealogy [ 24 ]. SWGDAM has provided an overview of IGG that emphasizes the approach being used only after a regular STR profile search of a law enforcement DNA database fails to produce any investigative leads [ 102 ]. Policy and practical implications of IGG have been explored in Australia [ 103 ] and within the UK as part of probing the perceptions of 45 professional and public stakeholders [ 104 , 105 ].

Four misconceptions about IGG were examined by several members of the SWGDAM group: (1) when law enforcement conducts IGG in a genetic genealogy database, they are given special access to participants' SNP profiles, (2) law enforcement will arrest a genetic genealogy database participant's relatives based on the genetic information the participant provided to the database, (3) IGG necessarily involves collecting and testing DNA samples from a larger number of innocent persons than would be the case if IGG were not used in the investigation, and (4) IGG is or soon will be ubiquitous because there are no barriers to IGG that limit the cases in which it can be conducted [ 106 ].

In May 2021, the state of Maryland passed the first law in the United States and in the world that regulates law enforcement's use of DTC genetic data to investigate crimes. A policy forum article in Science explained how this new law provides a model for others in this area [ 107 ]. Six important features were described: (1) requiring judicial authorization for the initiation of an IGG search, (2) affirming individual control over the investigative use of one's genetic data, (3) establishing strong protections for third parties who are not suspects in the case, (4) ensuring that IGG is available to prove either guilt or innocence, (5) imposing consequences and fines for violations, and (6) requiring annual public reporting and review to enable informed oversight of IGG methods. However, as of September 2022, these regulations have not been implemented apparently due to lack of resources with these unfunded requirements. 37

Efforts have been made to raise awareness among defense attorneys about how IGG searches can potentially invade people's privacy in unique ways [ 108 ]. Important perspectives on ethical, legal, and social issues have been offered along with directions for future research [ 109 ]. These concerns about data privacy, public trust, proficiency and agency trust, and accountability have led to a call for standards and certification of IGG to address issues raised by privacy scholars, law enforcement agencies, and traditional genealogists [ 110 , 111 ] and for an ethical and privacy assessment framework covering transparency, access criteria, quality assurance, and proportionality [ 112 ].

2.2.4. Genetic privacy and ethical concerns

Two important topics are considered in this section: (1) do the genetic markers used in traditional forensic DNA typing reveal more than identity and therefore potentially impact privacy of the individuals tested? and (2) are samples collected and tested according to ethical principles?

Forensic DNA databases utilize STR markers that were intentionally selected to avoid phenotypic associations. An extensive review of the literature examined 107 articles associating a forensic STR with some genetic trait and found “no demonstration of forensic STR variants directly causing or predicting disease” [ 113 ]. A study of the potential association of 15 STRs and 3 facial characteristics on 721 unrelated Han Chinese individuals also found “scarcely any association between [the] STRs with studied facial characteristics” [ 114 ].

In 2021, the American Type Culture Collection (ATCC) published a standard for authentication of human cell lines using DNA profiling with the 13 CODIS STR markers [ 115 ]. This use of forensic STR markers for biospecimen authentication led a bioethicist and a law professor to write a policy forum article in Science titled “Get law enforcement out of biospecimen authentication” [ 116 ]. The authors of this policy forum believe that using the same genetic markers could potentially: (1) undermine efforts to recruit research participants from historically marginalized and excluded groups that are underrepresented in research, (2) risk drawing law enforcement interest in gaining access to these research data, and (3) impose additional potential harms on already vulnerable populations, particularly children. Instead they advocate for using non-CODIS STRs or a new SNP assay to distinguish biospecimens in repositories, something done recently at the Coriell Institute for Medical Research with six new STR markers [ 117 ]. A responsive letter to the editor regarding this policy forum article expressed that “their proposal could potentially create artificial silos between genomic data in the justice system and in biomedical research, making it inefficient and ultimately counterproductive” [ 118 ]. The authors of the original article responded that “the risk of attracting law enforcement interest to research data increases when the data are available in a recognizable way” [ 119 ].

Modern scientific research seeks to protect the dignity, rights, and welfare of research participants by following ethical requirements. Six forensic science journals over the time period of 2010–2019 were examined for their reporting of ethical approval and informed consent in original research using human or animal subjects [ 120 ]. These journals were Forensic Science International: Genetics , Science & Justice , Journal of Forensic and Legal Medicine , the Australian Journal of Forensic Sciences , Forensic Science International , and the International Journal of Legal Medicine . A total of 3010 studies that described research on human or animal subjects and/or samples were selected from these journals with only 1079 articles (36%) reporting that they had obtained ethical approval and 527 articles (18%) stating that informed consent was sought either by written or verbal agreement. The authors of this study noted that reported compliance with ethical guidelines in forensic science research and publication was below what is considered minimal reporting rates in biomedical research and encouraged widespread adoption of the 2020 guidelines described below [ 120 ].

Guidelines and recommendations for ethnical research on genetics and genomics of biological material were jointly adopted and published in Forensic Science International: Genetics [ 121 ] and Forensic Science International: Reports [ 122 ]. These guidelines utilize the following principles as prerequisites for publication in these two journals as well as the Forensic Science International: Genetics Supplement Series : (1) general ethics principles that are regulated by national boards and represent widely signed international agreements, (2) universal declarations that require implementations in state members, such as the World Medical Association Declaration of Helsinki biomedical research on human subjects, and (3) universal declarations and principles drafted by independent organizations that have been widely adopted by the scientific community. This includes the U.S. Federal Policy for the Protection of Human Subjects (“Common Rule”) that was revised in 2017 (with a compliance date delayed to January 21, 2019). 38

Submitted manuscripts must provide the following supporting documentation to demonstrate compliance with the publication guidelines: (1) ethical approval in the country of [sample] collection by the appropriate local ethical committee or institutional review board, (2) ethical approval in the country of experimental work according to local legislation; if material collection and experimentation are conducted in different countries, both (1) and (2) are required, (3) template of consent forms in the case of human material as approved by the relevant ethical committee, and (4) approved export/import permits as applicable. Authors must declare in their submitted manuscript that these guidelines have been strictly followed [ 121 , 122 ].

Forensic genetic frequency databases, such as the Y-chromosome Haplotype Reference Database (YHRD), have been challenged over the ethics of DNA holdings, specifically of samples originating from the minority Muslim Uyghur population in western China [ 123 , 124 ]. A survey of U.S. state policies on potential law enforcement access to newborn screening samples found that nearly one-third of states permit these samples or their related data to be disclosed to or used by law enforcement and more than 25% of states have no discernible policy in place regarding law enforcement access [ 125 ].

A framework for ethical conduct of forensic scientists as “lived practice” has been proposed, and three case studies were discussed in terms of decision-making processes involving forensic DNA phenotyping and biographical ancestry testing, investigative genetic genealogy, and forensic epigenetics [ 126 ]. An ethos for forensic genetics involving the values of integrity, trustworthiness, and effectiveness has likewise been described [ 127 ].

2.2.5. Sexual assault kit testing

Unsubmitted or untested sexual assault kits (SAKs) may exist in police or laboratory evidence lockers for many years leading to rape kit backlogs that can spark community outrage when discovered. A number of articles have been published in the past three years describing success rates with examining SAKs and the policies surrounding them. For example, an evaluation of 3422 unsubmitted SAKs in Michigan found 1239 that produced a DNA profile eligible for upload into CODIS with 585 yielding a CODIS hit [ 128 ]. In addition, results from a groping and sexual assault case were presented to support the expansion of touch DNA evidence in these types of cases [ 129 ].

To assess success rates in their jurisdiction, the Houston Police Department randomly selected 491 cases of over 6500 previously unsubmitted sexual assault kits [ 130 ]. Of these, 336 cases (68%; 336/491) screened positive for biological evidence; a DNA profile was developed in 270 cases (55%; 270/491) with 213 (43%; 213/491) uploaded to CODIS; and 104 (21% total; 104/491 or 49% of uploaded profiles; 104/213) resulted in a CODIS hit. The statute of limitation had expired in 44% of these CODIS-hit cases, which prohibited arrests and prosecution. Victims were unwilling to participate in a follow-up investigation in another 25% of these cases. When the data were compiled for the publication, charges had been filed in only one CODIS-hit case [ 130 ].

Sexual assault cases can be difficult to prosecute as victims may be re-traumatized when a cold case is reopened. The authors of one study shared: “A key to successful pursuit of cold case sexual assaults is to have a well-crafted victim-notification plan and a victim advocate as part of the investigative team” [ 131 ]. Interviews with eight assistant district attorneys provided important prosecutors’ perspectives on SAK cases, the development of narratives to explain the evidence in a case, and the decision on whether a case should be pursued or what further investigative activities may be needed [ 132 ]. The authors concluded: “Our findings suggest that forensic evidence does not magically lead to criminal justice outcomes by itself, but must be used thoughtfully in conjunction with other evidence as part of a well-considered strategy of investigation and prosecution” [ 132 ].

Discussing a data set from Denver, Colorado where 1200 sexual assault cold cases with testable DNA samples were examined and 600 cases were processed through the laboratory resulting in 97 CODIS hits, 55 arrests and court filings, and 48 convictions, the authors conclude that the cost of the Denver cold case sexual assault program was worth the investment [ 131 ].

From December 2015 to July 2018, the Palm Beach County Sheriff's Office (Florida, USA) researched more than 5500 cases and evaluated evidence from previously untested sexual assault kits spanning a 43-year period at a cost of over $1 million. Of the 1558 sexual assaults examined, there were 686 cases (44%; 686/1558) with CODIS-eligible profiles, 261 CODIS hits, and 5 arrests when the article was written in mid-2019 [ 133 ]. The Palm Beach County Sheriff's Office also helped develop a backlog reduction effort through creating a biological processing laboratory within the Boca Raton Police Services Department [ 134 ]. With this joint effort from 2016 to 2018, the total average turnaround time decreased from 30 days to under 20 days with the 3489 DNA profiles entered into CODIS resulting in 1254 associations and 965 investigations aided. Important takeaway lessons include the value of (1) engaging legal counsel early to outline necessary legal procedures and the timeline, (2) bringing all stakeholders “to the table” early to discuss expectations, as well as legal and operational responsibilities, and (3) creating a realistic timeline with a comprehensive memorandum of understanding so all parties have agreed to their roles and responsibilities [ 134 ].

From 275 previously untested sexual assault kits submitted for DNA testing in one region of Central Brazil, a total of 176 profiles were uploaded to their DNA database resulting in 60 matches (34%; 60/176) and 32 assisted investigations (18%; 32/176) with information about the suspect identity or the connection of serial sexual assaults assigned to the same individual [ 135 ]. Another study from the same region of Brazil examined 2165 cases and noted that 13% (286/2165) had information regarding the victim-offender relationship with 63% (179/286) being stranger-perpetrated rapes and 37% (107/286) being non-stranger [ 136 ]. The authors then summarize: “Hits were detected only with stranger-perpetrated assaults ( n = 41), which reinforces that DNA databases are fundamental to investigate sexual crimes. Without DNA typing and DNA databases, probably these cases would never be solved” [ 136 ].

Given that laboratories have limited resources and need to prioritize their efforts, some business analytics have been applied to SAK testing. An analysis of the potential societal return on investment (ROI) for processing backlogged, untested SAKs reported a range of 10%–65% ROI depending on the volume of activity for the laboratory conducting the analysis [ 137 ]. An evaluation of data from 868 SAKs tested by the San Francisco Policy Department Criminalistics Laboratory during 2017–2019 found that machine learning algorithms outperformed forensic examiners in flagging potentially probative samples [ 138 ].

An examination of 5165 SAKs collected in Cuyahoga County (Ohio, USA) from 1993 through 2011 found 3099 with DNA of which 2127 produced a CODIS hit, with 803 investigations leading to an indictment and eventually 78 to trial along with 330 pleas [ 139 ]. The authors report a “cost savings to the community of $26.48 million after the inclusion of tangible and intangible costs of future sexual assaults averted through convictions” and advocate for “the cost-effectiveness of investigating no CODIS hit cases and support an ‘investigate all’ approach” [ 139 ]. Likewise an assessment of 900 previously-untested SAKs from Detroit (Michigan, USA) found that “few of the tested variables were significant predictors of CODIS hit rate” and “testing all previously-unsubmitted kits may generate information that is useful to the criminal justice system, while also potentially addressing the institutional betrayal victims experienced when their kits were ignored” [ 140 ].

A group in the Philippines described an integrated system to improve their SAK processing [ 141 ]. With an optimized workflow in Montreal, Canada, SAK processing median turnaround time decreased from 140 days to 45 days with a foreign DNA profile being obtained in 44% of cases [ 142 ]. In addition, this group examined casework data to guide resource allocation through identifying the likelihood of specific types of cases and samples yielding foreign biological material [ 142 ]. Decision trees and logistic regression models were also used to try and predict whether or not SAKs will yield a CODIS-eligible DNA profile [ 143 ]. Finally, direct PCR and rapid DNA approaches to streamline SAK testing were reviewed [ 144 ].

2.3. Forensic biology and body fluid identification

The basic workflow for biological samples in forensic examinations typically involves a visual examination of the evidence, a presumptive and/or confirmatory test for a suspected body fluid (e.g., the amylase assay for saliva), and DNA analysis and interpretation [ 145 ]. Body fluid identification (BFID), in particular with blood, saliva, semen, or vaginal fluid stains, provides valuable evidence in many investigations that can aid in the resolution of a crime [ 146 ]. Many of these BFID tests are presumptive and not nearly as sensitive as modern DNA tests meaning that “obtaining a DNA profile without being able to associate [it] with a body fluid is an increasingly regular occurrence” and “it is necessary and important, especially in the eyes of the law, to be able to say which body fluid that the DNA profile was obtained from” [ 147 ].

A number of approaches are being taken to improve the sensitivity and specificity of BFID in recent years including DNA methylation [ [148] , [149] , [150] , [151] , [152] , [153] , [154] , [155] , [156] , [157] , [158] , [159] , [160] , [161] ], messenger RNA (mRNA) [ [162] , [163] , [164] , [165] , [166] ], microRNA (miRNA) [ 167 ], protein mass spectrometry for seminal fluid detection [ 168 ], and microbiome analysis [ 169 , 170 ]. Although many new techniques are being described in the scientific literature, traditional methods for semen identification are still widely used in regular forensic casework [ 171 ].

When using RNA assays, DNA and RNA are co-extracted from examined samples [ 172 , 173 ]. Some tests may only distinguish between two possible body fluids, such as saliva and vaginal fluid [ 174 ], while other tests may attempt to distinguish six forensically relevant body fluids – vaginal fluid, seminal fluids, sperm cells, saliva, menstrual blood, and peripheral blood – although not always as clearly as desired [ 175 ]. BFID assays must also cope with mixed body fluids [ 176 ].

2.4. DNA collection and extraction

The process of obtaining a DNA profile begins with collecting a biological sample and extracting DNA from it. A review of recent trends and developments in forensic DNA extraction focused on isolating male DNA in sexual assault cases, using portable rapid DNA testing instruments, recovering DNA from difficult samples such as human remains, and bypassing DNA extraction altogether with direct PCR methods [ 177 ].

2.4.1. Touch evidence and fingerprint processing methods

Various studies have explored the compatibility of common fingerprint processing methods with DNA typing results [ [178] , [179] , [180] , [181] , [182] , [183] , [184] , [185] , [186] , [187] , [188] ]. For example, DNA recovery was explored after various steps in three different latent fingerprint processing methods – and fewer treatments were judged preferable with a 1,2-indanedione-zinc (IND/Zn) method appearing least harmful to downstream DNA analysis [ 187 ]. A different study found improved recovery of DNA from cigarette butts following latent fingerprint processing with 1,8-diazafluoren-9-one (DFO) compared to IND/Zn [ 179 ].

DNA losses were quantified with mock fingerprints deposited on four different surfaces to better understand DNA collection and extraction method performance [ 189 ]. The application of Diamond Dye has been shown to enable visualization of cells deposited on surfaces without interfering with subsequent PCR amplification and DNA typing [ [190] , [191] , [192] ].

It was possible to recover DNA profiles from clothing that someone touched for as little as 2 s [ 193 ]. DNA sampling success rates from car seats and steering wheels were studied [ 194 ] and recovery of DNA from vehicle surfaces using different swabs was explored [ 195 ]. In addition, the double-swab technique, where a wipe using a wet swab is followed by a wipe with a dry one, was revisited with an observation that for non-absorbing surfaces, the first web swab yielded 16 times more DNA than the second dry swab [ 196 ]. Swabs of cotton, flocked nylon, and foam reportedly provided equivalent DNA recoveries for smooth/non-absorbing surfaces, and an optimized swabbing technique involving the application of a 60-degree angle and rotating the swab during sampling improved DNA yields for cotton swabs [ 197 ].

2.4.2. Results from unfired and fired cartridge cases

Ammunition needs to be handled to load a weapon and thus DNA from the handler may be deposited onto the ammunition via touch [ 198 ]. Important progress has been made in recovering DNA from ammunition such as unfired cartridges or fired cartridge cases (FCCs) that may remain at a crime scene after a weapon has been fired. Trace quantities of DNA recovered from firearm or FCC surfaces has been used to try and link results to gun-related crimes.

A 2019 review of the literature regarding obtaining successful DNA results from ammunition examined collection techniques, extraction methodologies, and various amplification kits and conditions [ 199 ]. A direct PCR approach detected more STR alleles than methods using DNA extraction, and the authors noted that mixtures are commonly observed from gun surfaces, bullets, and cartridges in both controlled experimental conditions and from actual casework evidence and they encourage careful interpretation of these results [ 200 ]. The development of a crime scene FCC collector was combined with a new DNA recovery method that uses a rinse-and-swab technique [ 201 ].

Research studies and review articles have considered factors affecting DNA recovery from cartridge cases and the impact of metal surfaces on DNA recovery [ [202] , [203] , [204] , [205] , [206] , [207] , [208] , [209] ]. Recovery of mtDNA from unfired ammunition components has been assessed for sequence quality [ 210 ].

2.5. DNA typing

Following collection of DNA evidence and its extraction from biological samples, the typical typing process involves DNA quantitation, PCR amplification of STR markers, and STR typing using capillary electrophoresis. Direct PCR avoids the DNA extraction and quantitation steps, which can improve recovery of trace amounts of DNA [ 211 , 212 ]. Whole genome amplification prior to STR analysis has also been examined to aid recovery of degraded DNA [ 213 ] and to enable profiling of single sperm cells [ 214 ].

PCR amplification using STR typing kits can sometimes produce artifacts that impact DNA interpretation including missing (null) alleles [ 215 ], false tri-allelic patterns [ 216 ] or extra peaks when amplified in the presence of microbial DNA [ [217] , [218] , [219] ].

Applied Biosystems Genetic Analyzers have been the primary means of performing multi-colored capillary electrophoresis for many years [ 4 ]. First experiences with Promega's new Spectrum Compact CE System have recently been reported [ 220 ]. A number of new research and commercial STR kits have been introduced in recent years along with the publication of at least 24 validation studies ( Table 5 ). These validation studies typically follow guidelines outlined by the ENFSI DNA Working Group, 39 SWGDAM 40 , or a 2009 Chinese National Standard. 41

STR kits assessed with 24 published validation studies during 2019–2022.

A report on the first two years of submissions to the STRidER 42 (STRs for Identity ENFSI Reference) database for online allele frequencies revealed that 96% of the submitted 165 autosomal STR datasets generated by CE contained errors, showing the value of centralized quality control and data curation [ 245 ].

2.6. DNA interpretation at the source or sub-source level

The designation of STR alleles and genotypes of contributors in DNA mixtures are key aspects of DNA interpretation [ 246 , 247 ]. Electropherograms generated by CE instruments exhibit both STR alleles and artifacts that complicate data interpretation. Efforts are underway to understand and model instrumental artifacts [ [248] , [249] , [250] , [251] ] as well as biological artifacts of the PCR amplification process such as STR stutter products [ 252 , 253 ]. Machine learning approaches are being applied to classify artifacts versus alleles with the goal to eventually replace manual data interpretation with computer algorithms [ [254] , [255] , [256] , [257] ]. One such program, FaSTR DNA, enables potential artifact peaks from stutter, pull-up, and spikes to be filtered or flagged, and a developmental validation has been published examining 3403 profiles generated with seven different STR kits [ 258 ].

2.6.1. DNA mixture interpretation

Forensic evidence routinely contains contributions from multiple donors, which result in DNA mixtures. A number of approaches have been taken and advances made in DNA mixture interpretation [ 259 ]. These include probabilistic genotyping software [ 15 ], using genetic markers beyond traditional autosomal STR typing [ 260 ], or separating contributor cells and performing single-cell analysis [ [261] , [262] , [263] , [264] , [265] , [266] ].

In June 2021, the National Institute of Standards and Technology (NIST) released a draft report regarding the scientific foundations of DNA mixture interpretation [ 267 ]. This 250-page document described 16 principles that underpin DNA mixture interpretation, provided 25 key takeaways, and cited 528 references. NIST also began a Human Factors Expert Working Group on DNA Interpretation in February 2020 and plans to release a report with recommendations in 2023.

Assessment of the number of contributors (NoC) is a critical element of accurate DNA mixture interpretation. For example, the LRs relating to minor contributors can be reduced when the incorrect number of contributors is assumed [ 268 ]. Allele sharing among contributors to a mixture and masking of alleles due to STR stutter artifacts can lead to inaccurate NoC estimates based on simply counting the number of alleles at a locus. Different approaches and software programs have been used for NoC estimation [ [269] , [270] , [271] , [272] , [273] , [274] , [275] ]. Total allele count (TAC) distribution via TAC curves showed an improvement in manually estimating the number of contributors with complex mixtures [ 276 ]. Sequence analysis of STR loci expands the number of possible alleles compared to CE-based length measurements and thus can improve NoC estimates [ 277 ].

In the past three years, validation studies have been performed with a number of probabilistic genotyping software (PGS) systems including EuroForMix [ 278 ], DNAStatistX [ 279 , 280 ], TrueAllele [ 281 ], STRmix [ 282 ], Statistefix [ 283 ], Mixture Solution [ 284 ], Kongoh [ 285 ], and MaSTR [ 286 , 287 ]. Developers of EuroForMix, DNAStatistX, and STRmix provided a review of these systems [ 288 ]. Multi-laboratory assessments have been described [ 289 , 290 ] and likelihood ratios obtained from EuroForMix and STRmix compared [ [291] , [292] , [293] , [294] ]. With a growing literature in this area, there are many other articles that could have been cited.

2.7. DNA interpretation at the activity level

DNA interpretation at the source or sub-source level helps to answer the question of who deposited the cell material, whether attribution for the result can be made to a specific cell type (i.e., source level) or simply to the DNA if no attribution can be made to a specific cell type (i.e., sub-source level). Activity-level propositions seek to answer the question of how did an individual's cell material get there. Interpretation at the activity level is sometimes referred to as evaluative reporting [ 295 , 296 ].

In 2020, the ISFG DNA Commission [ 38 ] discussed the why, when, and how to carry out evaluative reporting given activity level propositions through providing examples of formulating these propositions. These Commission recommendations emphasize that reports using a likelihood ratio based on case-specific propositions and relevant conditioning information should highlight the assumptions being made and that “it is not valid to carry over a likelihood ratio from a low level, such as sub-source, to a higher level such as source or activity propositions … because the LRs given sub-source level propositions are often very high and LRs given activity level propositions will often be many orders of magnitude lower” [ 38 ]. Another recommendation specifies that “scientists must not give their opinion on what is the ‘most likely way of transfer’ (direct or indirect), as this would amount to giving an opinion on the activities and result in a prosecutor's fallacy (i.e., give the probability that X is true). The scientists' role is to assess the value of the results if each proposition is true in accordance with the likelihood ratio framework (the probability of the results if X is true and if Y is true)” [ 38 ] (emphasis in the original). This DNA Commission provided 11 recommendations and 4 considerations that should be studied carefully by those who implement activity-level DNA interpretation.

2.7.1. DNA transfer and persistence studies

To evaluate DNA findings given activity-level propositions it is important to understand the factors and variables that may impact DNA transfer, persistence, prevalence, and recovery (DNA-TPPR). These factors include history of contacting surfaces, biological material type, quantity and quality of DNA, dryness of biological material, manner and duration of contact, number and order of contacts, substrate type(s), time lapses and environment, and methods and thresholds used in the forensic DNA laboratory to generate the available data [ 297 ].

Three valuable review articles were published on this topic in 2019 [ 14 , 28 , 299 ]. Following a comprehensive January 2019 review that cited [ 298 ] references on DNA-TPPR [ 14 ], the same authors provided an update in November 2021 on recent progress towards meeting challenges and a synopsis of 144 relevant articles published between January 2018 and March 2021 [ 297 ]. While few studies provide the information needed to help assign probabilities of obtaining DNA results given specific sets of circumstances, progress includes use of Bayesian Networks [ 300 ] to identify variables for complex transfer scenarios [ 38 , [301] , [302] , [303] , [304] , [305] ] as well as development of an online database DNA-TrAC 43 for relevant research articles [ 299 ] and a structured knowledge base 44 with information to help practitioners interpret general transfer events at an activity level [ 306 ].

Forensic DNA pioneer Peter Gill emphasized that awareness of the limitations of DNA evidence is important for users of this data given that an increased sensitivity of modern DNA methods means that DNA may be recovered that is irrelevant to the crime under investigation [ 307 ]. An ISFG DNA Commission (see Section 1.2.5 ) emphasized that the strength of evidence associated with a DNA match at the sub-source level cannot be carried over to activity level propositions [ 38 ]. Structuring case details into propositions, assumptions, and undisputed case information has been encouraged [ 308 ].

Factors affecting variability of DNA recovery on firearms were studied with four realistic, casework-relevant handling scenarios along with results obtained including DNA quantities, number of contributors, and relative profile contributions for known and unknown contributors [ 309 ]. These studies found that sampling several smaller surfaces on a firearm and including the sampling location in the evaluation process can be helpful in assessing results given alternative activity-level propositions in gun-related crimes. The authors recommend that “further extensive, detailed and systematic DNA transfer studies are needed to acquire the knowledge required for reliable activity-level evaluations” [ 309 ].

Other recent studies on DNA-TPPR include examining prevalence and persistence of DNA or saliva from car drivers and passengers [ [310] , [311] , [312] ], evaluation of DNA from regularly-used knives after a brief use by someone else [ 313 ], studying the accumulation of endogenous and exogenous DNA on hands [ 314 ] and non-self-DNA on the neck [ 315 ], considering the potential of DNA transfer via work gloves [ 316 , 317 ] or during lock picking [ 318 ], and investigating whether DNA can be recovered from illicit drug capsules [ 319 , 320 ] or packaging [ 321 ] to identify those individuals preparing or handling the drugs.

Efforts have been made to estimate the quantity of DNA transferred in primary versus secondary transfer scenarios [ 322 ]. As quantities of DNA transferred can be highly variable and thought to be dependent on the so-called “shedder status” – how much DNA an individual exudes, several studies explored this topic [ [323] , [324] , [325] , [326] , [327] ]. Studies have also considered the level of DNA an individual transfers to untouched items in their immediate surroundings [ 328 ], the position and level of DNA transferred during digital sexual assault [ 329 ] or during various activities with worn upper garments [ 330 , 331 ], and the DNA composition on the surface of evidence bags pre- and post-exhibit examination [ 332 ]. Studies assessing background levels of male DNA on underpants worn by females [ 333 ] and background levels of DNA on flooring within houses [ 334 ] are providing important knowledge about the possibilities and probabilities of DNA transfer and persistence.

The authors of one study summarize some key points that could be extended to many other studies as words of caution: “From a wider trace DNA point of view, this study has demonstrated that the person who most recently handled an item may not be the major contributor and someone who handled an item for longer may still not be the major contributor if they remove more DNA than they deposit. The amount of DNA transferred and retained on an item is highly variable between individuals and even within the same individual between replicates” [ 320 ].

3. Emerging technologies, research studies, and other topics

New technologies to aid forensic DNA typing are constantly under development. This section explores recent activities with next-generation DNA sequencing, DNA phenotyping for estimating a sample donor's age, ancestry, and appearance, lineage markers, other markers and approaches, and non-human DNA and wildlife forensics, and is expected to be of value to researchers and those practitioners looking to future directions in the field.

3.1. Next-generation sequencing

Next-generation sequencing (NGS), also known as massively parallel sequencing (MPS) in the forensic DNA community, expands the measurement capabilities and information content of a DNA sample beyond the traditional length-based results with STR markers obtained with capillary electrophoresis (CE) methods. Additional genetic markers, such as single nucleotide polymorphisms (SNPs), microhaplotypes, and mitochondrial genome (mtGenome) sequence, may be analyzed along with the full sequence of STR alleles. This higher information content per sample opens up new potential applications such as phenotyping of externally visible characteristics and biogeographical ancestry as described in review articles [ 335 , 336 ].

As mentioned in Section 1.2.1 , the NIJ Forensic Laboratory Needs Technology Working Group (FLN-TWG) published a 29-page implementation strategy on next-generation sequencing for DNA analysis in September 2021 [ 28 ]. This guide discusses how NGS works and its advantages and disadvantages, the various instrument platforms and commercial kits available with approximate costs, items to consider regarding facilities, data storage, and personnel training, and resources for implementing NGS technology. A total of 73% of 105 forensic DNA laboratories surveyed from 32 European countries already own an MPS platform or plan to acquire one in the next year or two and one-third of the survey participants already conduct MPS-based STR sequencing, identity, or ancestry SNP typing [ 337 ].

Validation studies have been described with the ForenSeq DNA Signature Prep kit and the MiSeq FGx system [ [338] , [339] , [340] ], with the Verogen ForenSeq Primer Mix B for phenotyping and biogeographical ancestry predictions [ 341 , 342 ], and for resizing reaction volumes with the ForenSeq DNA Signature Prep kit library preparation [ 343 ]. MPS sequence data showed excellent allele concordance with CE results for 31 autosomal STRs in the Precision ID GlobalFiler NGS STR Panel from 496 Spanish individuals [ 344 ] and from 22 autosomal STR loci in the PowerSeq 46GY panel with 247 Austrians [ 345 ].

STR flanking region sequence variation has been explored [ 346 ] and reports of population data and sequence variation were published for samples from India [ 347 ], France [ 348 ], China [ 349 , 350 ], Korea [ 351 ], Brazil [ 352 ], Tibet [ 353 ], and the United States [ 354 ].

In April 2019 the STRAND ( S hort T andem R epeat: A lign, N ame, D efine) Working Group was formalized [ 355 ] to consider several possible approaches to sequence-based STR nomenclature that have been proposed [ 356 , 357 ]. An overview of software options has been provided for analysis of forensic sequencing data [ 358 ]. Some recent published options include STRinNGS [ 359 ], STRait Razor [ 360 ], ArmedXpert tools MixtureAce and Mixture Interpretation to analyze MPS-STR data [ 361 ], and STRsearch for targeted profiling of STRs in MPS data [ 362 ]. To aid interpretation of MPS-STR data, sensitivity studies were performed with single-source samples and sequence data analyzed by DNA quantity and method used [ 363 ]. A procedure has been described to address calculation of match probabilities when results are generated using MPS kits with different trim sites than those present in the relevant population frequency database [ 364 ]. Performance of different MPS kits, markers, or methods can be compared for accuracy and precision using the Levenshtein distance metric [ 365 ].

Novel MPS STR and SNP panels developed in recent years include IdPrism [ 366 ], a QIAGEN 140-locus SNP panel [ 367 ], the 21plex monSTR identity panel [ 368 ], a 42plex STR NGS panel to assist with kinship analysis [ 369 ], the 5422 marker FORCE (FORensic Capture Enrichment) panel [ 370 ], a forensic panel with 186 SNPs and 123 STRs [ 371 ], the SifaMPS panel for targeting 87 STRs and 294 SNPs [ 372 ], a 1245 SNP panel [ 373 ], 90 STRs and 100 SNPs for application with kinship cases [ 374 ], an adaption of the SNPforID 52plex panel to MPS [ 375 ], 448plex SNP panel [ 376 ], a 133plex panel with 52 autosomal and 81 Y-chromosome STRs [ 377 ], and a forensic identification multiplex with 1270 tri-allelic SNPs involving 1241 autosomal and 29 X-chromosome markers [ 378 ]. The 124 SNPs in the Precision ID Identity Panel were examined in a central Indian population [ 379 ] and human leukocyte antigen (HLA) alleles used in the early 1990s were revisited with MPS capability [ [380] , [381] , [382] ].

MPS methods have demonstrated utility with compromised samples [ [383] , [384] , [385] , [386] , [387] , [388] ] and mixture interpretation [ [389] , [390] , [391] , [392] , [393] , [394] , [395] ]. Microhaplotype assays have also been developed to assist with DNA mixture deconvolution [ 396 , 397 ]. Collaborative studies have explored variability with laboratory performance using MPS methods [ 398 , 399 ]. Population structure [ 400 ] and linkage and linkage disequilibrium [ 401 ] were examined among the markers in forensic MPS panels.

A review of transcriptome analysis using MPS discussed efforts with body fluid and tissue identification, determination of the time since deposition of stains and the age of donors, the estimation of post-mortem interval, and assistance to post-mortem death investigations [ 402 ]. The potential for MPS methods to assist with environmental trace analysis was reviewed in terms of forensic soil analysis, forensic botany, and human identification utilizing the skin microbiome [ 403 ]. The possibility of non-invasive prenatal paternity testing using cell-free fetal DNA from maternal plasma was explored with the Precision ID Identity Panel [ 404 ] and the ForenSeq DNA Signature Prep Kit [ 405 ]. Pairwise kinship analysis was also examined using the ForenSeq DNA Signature Prep Kit and multi-generational family pedigrees [ 406 , 407 ]. Nanopore sequencing has also been explored for sequencing STR and SNP markers [ [408] , [409] , [410] , [411] , [412] , [413] , [414] , [415] , [416] ].

3.2. DNA phenotyping (ancestry, appearance, age)

Continuing research into the genetic components of biogeographic ancestry, appearance, and age predictions have improved forensic DNA phenotyping capabilities [ 417 ]. These forensic innovations may sometimes impact public expectations [ 418 ]. The investigation in a murder case was assisted using information from forensic DNA phenotyping that predicted eye, hair, and skin color of an unknown suspect with the HIrisPlex-S system involving targeted massively parallel sequencing [ 419 ].

The VISAGE ( Vis ible A ttributes Through Ge nomics) Consortium, which consists of 13 partners from academic, police, and justice institutions in 8 European countries, has established new scientific knowledge and developed and tested prototype tools for DNA analysis and statistical interpretation as well as conducted education for stakeholders. In the 2019 to 2022 time window of this review, this concerted effort produced 45 one review article [ 417 ], 22 original research publications [ 337 , [420] , [421] , [422] , [423] , [424] , [425] , [426] , [427] , [428] , [429] , [430] , [431] , [432] , [433] , [434] , [435] , [436] , [437] , [438] , [439] , [440] ], and three reports [ [441] , [442] , [443] ].

DNA phenotyping is currently an active area of research, and numerous activities and publications exist beyond the VISAGE articles noted here. Another 137 articles have appeared in the literature in the past three years on biogeographical ancestry, appearance (primarily hair color, eye color, and skin color), and biological age predictions (typically utilizing DNA methylation) (see Supplemental File ).

3.3. Lineage markers (Y-chromosome, mtDNA, X-chromosome)

Lineage markers consist of Y-chromosome, mitochondrial DNA, and X-chromosome genetic information that may be inherited from just one parent without the regular recombination that occurs with autosomal DNA markers. Research in terms of new markers, assays, and population studies continue to be published for these lineage markers.

3.3.1. Y-chromosome

Several recent review articles were published on forensic applications of Y-chromosome testing [ [444] , [445] , [446] ]. As discussed previously in Section 1.2 , an ISFG DNA Commission summarized the state of the field with Y-STR interpretation [ 39 ]. Rapidly mutating Y-STR loci can be used to differentiate closely related males [ [447] , [448] , [449] ]. New statistical approaches to assessing evidence with Y-chromosome information have been described [ 450 , 451 ]. Four commercial Y-STR multiplexes were compared with the NIST 1032 U S. population sample set and the allele and haplotype diversities explored with length-based versus sequence-based information [ 452 ].

A number of Y-STR typing systems have been described along with validation studies, such as a 36plex [ 453 ], a 41plex [ 454 ], a 29plex [ 455 ], a 17plex [ 456 ], a 24plex [ 457 ], the Microreader 40Y ID System [ 458 ], the 24 Y-STRs in the AGCU Y SUPP STR kit [ 459 ], the DNATyper Y26 PCR amplification kit [ 460 ], a multiplex with 12 multicopy Y-STR loci [ 461 ], the Yfiler Platinum PCR Amplification Kit [ 462 ], a 45plex [ 463 ], the Microreader 29Y Prime ID system [ 464 ], an assay with 30 slow and moderate mutation Y-STR markers [ 465 ], the 17plex Microreader RM-Y ID System [ 466 ], and a 26plex for rapidly mutating Y-STRs [ 467 ]. A machine learning program predicted Y haplogroups using two Y-STR multiplexes with 32 Y-STRs [ 468 ].

Deletions and duplications with 42 Y-STR were reported in a sample of 1420 unrelated males and 1160 father-son pairs from a Chinese Han population [ 469 ]. Using Y-STR allele sequences has enabled locating parallel mutations in deep-rooting family pedigrees [ 470 ]. The surname match frequency with Y-chromosome haplotypes was explored using 2401 males genotyped for 46 Y-STRs and 183 Y-SNPs [ 471 ]. In the Y-chromosome's role as a valuable kinship indicator to assist in genetic genealogy and forensic research, models to improve prediction of the time to the most recent common paternal ancestor have been studied with 46 Y-STRs and 1120 biologically related genealogical pairs [ 472 ]. A massively parallel sequencing tool was developed to analyze 859 Y-SNPs to infer 640 Y haplogroups [ 473 ]. Another MPS tool, the CSYseq panel, targeted 15,611 Y-SNPs to categorize 1443 Y-sub-haplogroup lineages worldwide along with 202 Y-STRs including 81 slow, 68 moderate, 27 fast, and 26 rapidly mutating Y-STRs to individualize close paternal relatives [ 474 ].

3.3.2. Mitochondrial DNA

Mitochondrial DNA (mtDNA), which is maternally inherited with a high copy number per cell, can aid human identification, missing persons investigations, and challenging forensic specimens containing low quantities of nuclear DNA such as hair shafts [ [475] , [476] , [477] ]. Validation studies have been published using traditional Sanger sequencing [ 478 ] and next-generation sequencing [ [479] , [480] , [481] ]. Illumina and Thermo Fisher now provide mtDNA whole genome NGS assays [ [482] , [483] , [484] , [485] ]. Many mtDNA population data sets were published in the past three years including high-quality data from U.S. populations [ 486 ]. The suitability of current mtDNA interpretation guidelines for whole mtDNA genome (mtGenome) comparisons has been evaluated [ 487 ].

NGS methods have increased sensitivity of mtDNA heteroplasmy detection [ 488 , 489 ], which can influence the ability to connect buccal reference samples and rootless hairs from the same individual [ 490 , 491 ]. Twelve polymerases were compared in terms of mtDNA amplification yields from challenging hairs – with KAPA HiFi HotStart and PrimeSTR HS outperforming AmpliTaq Gold DNA polymerase that is widely used in forensic laboratories [ 492 ]. Multiple studies and review articles have discussed distinguishing mtDNA from nuclear DNA elements of mtDNA (NUMTs) that have been inserted into our nuclear DNA [ [493] , [494] , [495] , [496] ].

NGS sequencing of the mtGenome has permitted improved resolution of the most common West Eurasian mtDNA control region haplotype [ 497 ]. Phylogenetic alignment and haplogroup classification have continued to be refined with new sequence information [ 498 ], and new assays have been developed to aid haplogroup classification [ 499 ]. Concerns over potential paternal inheritance of mtDNA have also been addressed [ 500 , 501 ].

3.3.3. X-chromosome

A 20-year review of X-chromosome use in forensic genetics examined the number and types of markers available, an overview of worldwide population data, the use of X-chromosome markers in complex kinship testing, mutation studies, current weaknesses, and future prospects [ 502 ]. One example of the forensic application of X-chromosome markers include use in relationship testing cases involving suspicion of incest or paternity without a maternal sample for comparison [ 503 ]. Four new X-STR multiplex assays were described along with validation studies including a 19plex [ 504 ], a 16plex [ 505 ], another 19plex – the Microreader 19X Direct ID System [ 506 ], and an 18plex named TYPER-X19 multiplex assay [ 507 ]. A collaborative study examined paternal and maternal mutations in X-STR markers [ 508 ]. A software program for performing population statistics on X-STR data was introduced [ 509 ] and sequence-based U.S. population data described for 7 X-STR loci [ 510 ].

3.4. New markers and approaches (microhaplotypes, InDels, proteomics, human microbiome)

In this section on new markers and approaches, publications related to microhaplotypes and insertion/deletion (InDel, or DIP for deletion insertion polymorphisms) markers are reviewed along with proteomic and microbiome approaches to supplement standard human DNA typing methods.

3.4.1. Microhaplotypes

Microhaplotype (MH) markers consist of multiple SNPs in close proximity (e.g., typically <200 bp or <300 bp) that can be simultaneously genotyped with each DNA sequence read using NGS. Two or more linked SNPs will define three or more haplotypes. Compared to STR markers, MHs do not have stutter artifacts (which complicate mixture interpretation), can be designed with shorter amplicon lengths in some cases (which benefits recovery of genetic information from degraded DNA samples), possess a higher degree of polymorphism compared to single SNP loci (which benefits discrimination power), and exhibit low mutation rates (which enables relationship testing and biogeographical ancestry inference). Thus, MH markers bring advantages to human identification, ancestry inference, kinship analysis, and mixture deconvolution to potentially assist missing person investigations, relationship testing, and forensic casework as discussed in several recent reviews [ 16 , 511 ]. A new database, MicroHapDB, has compiled information on over 400 published MH markers and frequency data from 26 global population groups [ 512 ].

A number of MH panels have been described [ [513] , [514] , [515] , [516] , [517] , [518] , [519] ]. Population data has been collected from a number of sources around the world including four U.S. population groups examined with a 74plex assay with 74 MH loci and 230 SNPs [ 520 ]. Various MH panels have been evaluated for effectiveness with kinship analysis [ [521] , [522] , [523] ]. Likewise the ability to detect minor contributors in DNA mixtures has been assessed [ [524] , [525] , [526] ].

3.4.2. InDel markers

InDel markers can be detected using a CE-based length analysis, and thus use instrumentation that forensic DNA laboratories already have. InDels can also be designed to amplify short DNA fragments (e.g., <125 bp) to help improve amplification success rates with low DNA quantity and/or quality. However, with only two possible alleles like SNPs, InDels are not as polymorphic as STRs and thus require more markers to obtain similar powers of discrimination as multi-allelic STR markers and do not work as well with mixed DNA samples. InDels possess a lower mutation rate than STRs and can be used as ancestry informative markers (AIMs) since allele frequencies may differ among geographically separated population groups.

Two commercial InDel kit exist: (1) Investigator DIPlex (QIAGEN, Hilden, Germany) with 30 InDels [ [527] , [528] , [529] , [530] , [531] ] and (2) InnoTyper 21 (InnoGenomics, New Orleans, Louisiana, USA) with 21 autosomal insertion-null (INNUL) markers [ [532] , [533] , [534] , [535] ]. In addition, a number of InDel assays have been published including a 32plex [ 536 ], a 35plex [ 537 ], a 38plex [ 538 ], a 39plex with AIMs [ 539 ], a 43plex [ 540 ], a 57plex [ 541 ], a 60plex with 57 autosomal InDels, 2 Y-chromosome InDels, and amelogenin [ 542 ], a 32plex with X-chromosome InDels [ 543 ], and a 21plex with AIMs [ 544 ].

A multi-InDel marker is a specific DNA fragment with more than one InDel marker located tightly in the physical position that provides a microhaplotype [ 545 ]. Several multi-InDel assays have been published include a 12plex [ 546 ] and an 18plex [ 547 ].

3.4.3. Proteomics

Protein analysis, often through immunological assays, has traditionally been used to identify body fluids and tissues. With improvements in protein mass spectrometry in recent years, genetic variation can be observed in hair shafts via single amino acid polymorphisms. Detection of these genetically variant peptides (GVPs) can infer the presence of corresponding SNP alleles in the genome of the individual who is the source of the protein sample. A thorough review of forensic proteomics in 2021 cited 375 references [ 18 ]. Recent efforts in this area have focused on using GVPs to differentiate individuals through their human skin cells [ [548] , [549] , [550] ] or hair samples [ [551] , [552] , [553] , [554] , [555] , [556] , [557] , [558] , [559] ]. An algorithm has been proposed for calculating random match probabilities with GVP information [ 560 ].

3.4.4. Human microbiome

Microorganisms live in and on the human body, and efforts are underway to utilize the human microbiome for a variety of potential forensic applications [ 21 , [561] , [562] , [563] ]. There are also active efforts with analysis of microbiomes in the environment (e.g., soil or water samples), which could be classified under non-human DNA testing. Forensic microbiome research covers at least six areas: (1) individual identification, (2) tissue/body fluid identification, (3) geolocation, (4) time since stain deposition estimation, (5) forensic medicine, and (6) post-mortem interval (PMI) estimation. Biological, technical, and data issues have been raised and potential solutions explored in a recent review article [ 21 ]. For example, microbes on deceased individuals are being studied to estimate the postmortem interval [ 20 ] and postmortem skin microbiomes were found to be stable during repeated sampling up to 60 h postmortem [ 564 ].

Sequence analysis of 16S rRNA using NGS provides information on the microbiome community present in a tested sample [ 565 ]. The Forensic Microbiome Database 46 correlates publicly available 16S rRNA sequence data as a community resource. If the skin microbiome is extremely diverse among individuals, then the potential exists to associate the bacterial communities on an individual's skin with objects touched by this individual assuming that the bacteria originating from the donor's skin are deposited (i.e., transfer to and persist on the surface) and can be detected and interpreted.

Specific aspects of the microbiome (e.g., the bacterial community) may be able to provide details about the donor through bacterial profiling. For example, in one study correlations were observed between the bacterial profile and gender, ethnicity, diet type, and hand sanitizer used [ 566 ]. Another study with 30 individuals found that each person left behind microbial signatures that could be used to track interaction with various surfaces within a building, but the authors concluded “we believe the human microbiome, while having some potential value as a trace evidence marker for forensic analysis, is currently under-developed and unable to provide the level of security, specificity and accuracy required for a forensic tool” [ 565 ].

Direct and indirect transfer of microbiomes between individuals has been studied [ 567 , 568 ] along with identifying background microbiomes [ 569 ] and the possibility of transfer of microbiomes within a forensic laboratory setting [ 570 ]. Changes in four bacterial species in saliva stains were charted, showing that it was possible to correctly predict deposition time within one week in 80% of the stains [ 571 ]. The ability to detect sexual contact has been explored through using the microbiome of the pubic region [ [572] , [573] , [574] ]. The microbiomes on skin, saliva, vaginal fluid, and stool samples have been compared [ 575 ]. The stability, diversity, and individualization of the human skin virome was explored with 59 viral biomarkers being found that differed across the 42 individuals studied [ 576 ]. It will be interesting to see what the future holds and what other findings come from this active area of research.

3.5. Kinship analysis, human identification, and disaster victim identification

Kinship analysis, which uses genetic markers and statistics to evaluate the potential for specific biological relationships, is important for parentage testing, disaster victim identification (DVI), and human identification of remains that may be recovered in missing person cases. New open-source software programs have been described that can assist with kinship analysis [ 577 , 578 ].

A potential biological relationship is commonly evaluated using a likelihood ratio (LR) by comparing the likelihoods of observing the genetic data given two alternative hypotheses, such as (1) an individual is related to another individual in a defined relationship versus (2) the two individuals not related. Higher LR values indicate stronger support with the genetic data if the proposed relationship is true. Multiple factors influence LR kinship calculations including the specific hypotheses, the genetic markers examined, the allele frequencies of the relevant population(s), the co-ancestry coefficient applied, and approaches to address potential mutations. STR genotypes were reported for 11 population groups used by the FBI Laboratory [ 579 ]. The status quo has been challenged in recent articles regarding how hypotheses are commonly established [ 580 ] and whether race-specific U.S. population databases should be used for allele frequency calculations [ 581 ].

Depending on the relationship being explored, information can be optimized through genetic information from additional known relatives or through collecting results at more loci [ 582 ]. Potential error rates have been modeled with the observation that false negatives, which occur when related individuals are misinterpreted as being unrelated, are more common than false positives, where unrelated people are interpreted as being related [ 583 ]. While LRs are generally reliable in detecting or confirming parent/child pairs, limitations of kinship determinations exist (e.g., distinguishing siblings from half-siblings) when using STR data [ 584 ].

Pairwise comparisons have been studied in forensic kinship analysis [ [585] , [586] , [587] ]. The effectiveness of 40 STRs plus 91 SNPs was shown to be better than 27 STRs and 91 SNPs or 40 STRs alone [ 588 ]. Only a minor increase in LRs was observed when taking NGS-generated allele sequence variation rather than fragment length allele variation [ 589 ]. The statistical power of exclusion and inclusion can be used to prioritize family members selected for testing in resolving missing person cases [ 590 ]. A strategy for making decisions when facing low statistical power in missing person and DVI cases was published [ 591 ].

The most challenging kinship cases involve efforts to separate pairs of individuals who are typically thought to be genetically indistinguishable (i.e., monozygotic twins) or distant relatives (e.g., fourth cousins) where there is an increased uncertainty in the possible relationship. In some situations, somatic mutations may permit distinguishing monozygotic twins following whole genome sequencing – and this approach was successful in four of six cases reported recently [ 19 ]. The probative value of NGS data for distinguishing monozygotic twins was explored [ 592 ]. A unique case of heteropaternal twinning was reported where opposite-sex twins apparently had different fathers [ 593 ]. An impressive effort in kinship analysis using direct-to-consumer genetic genealogy information from 56 living descendants of multiple genealogical lineages helped resolve a contested paternity case from over a century and a half ago to identify the biological father of Josephine Lyon [ 594 ].

Techniques for identification of human remains continue to improve particularly with the capabilities of NGS and hybridization capture [ 595 ] and ancient DNA extraction protocols [ 596 , 597 ]. Studies have reported variation in skeletal DNA preservation [ 598 ] and retrospectively considered success rates with compromised human remains [ 599 ].

A simulated airplane crash enabled six forensic laboratories in Switzerland to gain valuable DVI experience with kinship cases of varying complexity [ 600 ]. The ISFG Spanish-Portuguese Speaking Working Group likewise conducted a DVI collaborative exercise with a simulated airplane crash to explore fragment re-associations, victim identification through kinship analysis, coping with related victims, handling mutations or insufficient number of family references, working in a Bayesian framework, and the correct use of DVI software [ 601 ]. Other groups have explored the capability of a particular software tool [ 602 ] or implemented rapid DNA analysis to accelerate victim identification [ 603 ]. The International Commission on Missing Persons (ICMP) has gained considerable experience with DNA extraction and STR amplification from degraded skeletal remains and kinship matching procedures in large databases [ 604 ]. To supplement the INTERPOL DVI Guide, 47 some lessons learned and experienced-based recommendations for DVI operations have recently been provided [ 605 ].

3.6. Non-human DNA testing and wildlife forensics

Non-human biological evidence may inform criminal investigations when animals or plants are victims or perpetrators of crime or the presence of specific material, such as cat or dog hair, may contribute to reconstructing events at a crime scene. Non-human DNA testing includes wildlife forensics and domestic animal species as well as forensic botany and has many commonalities and some important differences compared to human DNA testing [ [606] , [607] , [608] , [609] , [610] ]. Pollen analysis can assist criminal investigations [ 611 , 612 ]. The potential for and the barriers associated with the wider application of forensic botany in civil proceedings and criminal cases have been examined [ 613 , 614 ].

Mammalian species identification can assist in determining the origins of non-human biological material found at crime scenes through narrowing the range of possibilities [ 615 ]. New sequencing methods have been developed to assist species identification [ 616 ]. A multiplex PCR assay was developed to simultaneously identify 22 mammalian species (alpaca, Asiatic black bear, Bactrian camel, brown rat, cat, cow, common raccoon, dog, European rabbit, goat, horse, house mouse, human, Japanese badger, Japanese wild boar, masked palm civet, pig, raccoon dog, red fox, sheep, Siberian weasel, and sika deer) and four poultry species (chicken, domestic turkey, Japanese quail, and mallard) [ 617 ]. A number of other species identification assays have also been reported [ [618] , [619] , [620] ].

An important effort for harmonizing canine DNA analysis is an ISFG working group known as the Canine DNA Profiling Group, or CaDNAP. 48 The CaDNAP group published an analysis of 13 STR markers in 1184 dogs from Germany, Austria, and Switzerland [ 621 ]. Six traits for predicting visible characteristics in dogs, namely coat color, coat pattern, coat structure, body size, ear shape, and tail length, were explored with 15 SNPs and six InDel markers [ 622 ]. Canine breed classification and skeletal phenotype prediction has been explored using various genetic markers [ 623 ]. A novel assay using a feline leukemia virus was developed to demonstrate that a contested bobcat was not a domestic cat hybrid [ 624 ] and a core panel of 101 SNP markers was selected for domestic cat parentage verification and identification [ 625 ].

DNA tests have been developed to assist with illegal trafficking investigations involving elephant ivory seizures [ 626 ], falcons [ 627 ], and precious coral material [ 628 ]. Accuracy in animal forensic genetic testing was explored with interlaboratory assessments performed in 2016 and 2018 [ 629 ]. A collaborative exercise conducted in 2020 and 2021 by the ISFG Italian Speaking Working Group examined performance across 21 laboratories with a 13-locus STR marker test for Cannabis sativa [ 630 ]. A molecular approach was explored to distinguish drug-type versus fiber-type hemp varieties [ 631 ].

Acknowledgments and disclaimer

I am grateful to Dominique Saint-Dizier from the French National Scientific Police for the invitation and opportunity to conduct this review and for the support of my supervisor, Shyam Sunder, for granting the time to work on this extensive review. Input and suggestions on this manuscript by Todd Bille, Thomas Callaghan, Kevin Kiesler, François-Xavier Laurent, Robert Ramotowski, Kathy Sharpless, and Robert Thompson are greatly appreciated. Certain commercial entities, equipment, or materials may be identified in this document in order to describe an experimental procedure or concept adequately. Such identification is not intended to imply recommendation or endorsement by the National Institute of Standards and Technology, nor is it intended to imply that the entities, materials, or equipment are necessarily the best available for the purpose.

1 https://www.sciencedirect.com/journal/forensic-science-international-genetics/special-issue/10TSDS4360H .

2 https://www.mdpi.com/journal/genes/special_issues/Forensic_Genetic .

3 https://www.mdpi.com/journal/genes/special_issues/forensic_mitochondrial_genomics .

4 https://www.mdpi.com/journal/genes/special_issues/Advances_Forensic_Genetics .

5 https://www.mdpi.com/books/pdfdownload/book/5798 .

6 https://www.mdpi.com/journal/genes/special_issues/Bioinformatics_Forensic_Genetics .

7 https://www.mdpi.com/journal/genes/special_issues/genetics_anthropology .

8 https://www.mdpi.com/journal/genes/special_issues/Identification_of_Human_Remains .

9 https://www.mdpi.com/journal/genes/special_issues/Forensic_DNA_analysis .

10 https://www.mdpi.com/journal/genes/special_issues/Forensic_DNA_Mixture .

11 https://www.mdpi.com/journal/genes/special_issues/28FBA0G4DH .

12 See https://www.swgdam.org/ .

13 https://www.swgdam.org/publications .

14 https://www.fbi.gov/file-repository/rapid-dna-guide-january-2022.pdf/view .

15 https://www.fbi.gov/file-repository/non-codis-rapid-dna-best-practices-092419.pdf/view .

16 https://www.fbi.gov/file-repository/rapid-dna-testing-for-non-codis-uses-considerations-for-court-073120.pdf/view .

17 https://www.justice.gov/olp/uniform-language-testimony-and-reports .

18 https://forensiccoe.org/human_factors_forensic_science_sourcebook/ .

19 https://www.nist.gov/organization-scientific-area-committees-forensic-science .

20 https://www.nist.gov/organization-scientific-area-committees-forensic-science/human-forensic-biology-subcommittee .

21 https://www.nist.gov/topics/organization-scientific-area-committees-forensic-science/wildlife-forensics-subcommittee .

22 https://www.aafs.org/academy-standards-board .

23 https://www.nist.gov/organization-scientific-area-committees-forensic-science/osac-registry .

24 See https://www.nist.gov/organization-scientific-area-committees-forensic-science/human-forensic-biology-subcommittee .

25 https://lexicon.forensicosac.org/ .

26 https://www.nist.gov/osac/human-factors-validation-and-performance-testing-forensic-science .

27 https://www.nist.gov/organization-scientific-area-committees-forensic-science/osac-research-and-development-needs .

28 https://www.gov.uk/government/publications/forensic-science-providers-codes-of-practice-and-conduct-2021-issue-7 .

29 https://www.aabb.org/standards-accreditation/standards/relationship-testing-laboratories .

30 https://www.isfg.org/DNA+Commission .

31 Previously available rapid DNA systems included the RapidHIT 200 from IntegenX and MiDAS (Miniaturized integrated DNA Analysis System) from the Center for Applied NanoBioscience at the University of Arizona.

32 See https://le.fbi.gov/science-and-lab-resources/biometrics-and-fingerprints/codis/rapid-dna .

33 See https://www.interpol.int/How-we-work/Forensics/DNA .

34 See https://www.interpol.int/How-we-work/Forensics/I-Familia .

35 See https://le.fbi.gov/science-and-lab-resources/biometrics-and-fingerprints/codis#Familial-Searching .

36 See https://isogg.org/wiki/Autosomal_DNA_testing_comparison_chart .

37 See https://www.wmar2news.com/infocus/maryland-quietly-shelves-parts-of-genealogy-privacy-law .

38 See https://www.hhs.gov/ohrp/regulations-and-policy/regulations/finalized-revisions-common-rule/index.html .

39 See https://enfsi.eu/about-enfsi/structure/working-groups/dna/ .

40 See https://www.swgdam.org/publications .

41 See https://www.chinesestandard.net/PDF/English.aspx/GAT815-2009 .

42 See https://strider.online/ .

43 See https://bit.ly/2R4bFgL (DNA-TrAC).

44 See https://cieqfmweb.uqtr.ca/fmi/webd/OD_CIEQ_CRIMINALISTIQUE (Transfer Traces Activity DataBase).

45 See https://www.visage-h2020.eu/index.html#publications .

46 See http://fmd.jcvi.org/ .

47 See https://www.interpol.int/en/How-we-work/Forensics/Disaster-Victim-Identification-DVI .

48 See https://www.isfg.org/Working+Groups/CaDNAP .

Appendix A Supplementary data to this article can be found online at https://doi.org/10.1016/j.fsisyn.2022.100311 .

Appendix A. Supplementary data

The following is the supplementary data to this article:

New hope for cold cases due to breakthrough in forensic fingerprint research

Researchers have unveiled a groundbreaking method capable of detecting drug substances from fingerprints lifted from crime scenes, which could provide fresh insights into unsolved cases.

finger prints on a strip held by someone wearing blue gloves

Analytical scientists from Loughborough University have demonstrated for the first time that drug residue – namely the fast-acting sleeping pill Zolpidem, which has been linked to drug-facilitated sexual assault and drink spiking – can be detected on gel-lifted fingerprints.

Dr Jim Reynolds and Dr Ayoung Kim say the breakthrough could shed new light on cold cases and unsolved crimes as forensic gel lifters – which transfer prints onto a gelatine surface – are used globally by scenes of crimes officers to preserve and visualise fingerprints.

“This is the first time that analysis of gel-lifted prints for a drug substance has been accomplished and shows that lifted prints and other forensic marks can be interrogated for useful information”, says Dr Reynolds, the research lead.

“Since gel-lifted prints and marks can be stored for many years, the technique could be of real use in cold cases where additional information may prove useful to either link or exonerate a suspect to the investigation.

“Working with police forces and applying the method to cold case samples could help bring criminals to justice who may have thought they have got away with it.”

A number of tests exist which can detect drugs directly from fingerprints, but these face limitations. They can be destructive to the fingerprint, degrade drug residues, and are affected by environmental interferences.

It has long been speculated that gel-lifted prints contain valuable chemical information and could offer more accurate drug detection.

However, traditional techniques used to analyse the chemicals present in a sample have previously not been suitable for gel lifters. This is because they detect all chemicals present, including those that make up the gel, making it difficult to identify specific substances.

The method used by Dr Reynolds and Dr Kim, called sfPESI-MS, overcomes this issue using a rapid separation mechanism that distinguishes the drug substance from the background of the gel.

The process involves sampling the chemicals from the gel lifters into tiny liquid droplets. The chemicals extracted into the droplets are then ionized, which means they gain or lose electric charge depending on their chemical properties. The drug substance chemicals are more surface active than the chemicals originating from the gel, which enables them to be separated from the mixture.

This separation method enables the direct detection of a drug substance using mass spectrometry, a technique that identifies chemicals by measuring their molecular weight.

Published in the Drug Testing and Analysis journal, the researchers have successfully tested the technique using Zolpidem-laced fingerprints lifted from glass, metal, and paper surfaces in a laboratory setting.

They now hope to work with police forces to analyse stored gel-lifted prints and use the method to identify other substances.

Dr Reynolds said: “Zolpidem was the focus of our research, but the method could just as easily be applied to other drug substances a person may have been handling and could be applied to other chemicals such as explosives, gunshot residues, paints, and dyes.

“By linking chemical information to the fingerprint, we can identify the individual and link to the handling of an illicit substance which may prove useful in a prosecution.

“This could be useful to detect individuals who have been spiking drinks, for example, if the drug they are using gets onto their fingertips, then they will leave evidence at the scene.”

Dr Kim, who is the first author of the paper and completed the research as part of her PhD at Loughborough, added: “We would like to apply our method to real samples from criminal investigations, it would be good to know my PhD research has helped bring criminals to justice.”

The paper can be read in full on the journal website.

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Forensic science standards: NIST is accelerating the development and adoption of high-quality, technically sound forensic science standards by administering the Organization of Scientific Area Committees for Forensic Science (OSAC). OSAC-approved standards define minimum requirements, best practices and standard protocols that help ensure that the results of forensic analysis are reliable and reproducible.
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Black background. Photo of a penny. Next to the penny is a tiny amount of a white powder.

Safe, Efficient, Reliable: New Science in the Fight Against Killer Drugs

“A new drug might appear, then three or six months later it’s gone, replaced by something new,” said NIST chemist and program manager Marcela Najarro. “It’s a totally different ballgame than 10 or 15 years ago.”

To help with that issue and others, NIST researchers are giving law enforcement and public health experts new tools to combat fentanyl and other synthetic drugs.

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NIST Publishes Review of DNA Mixture Interpretation Methods

Spotlight: forensics videos and app for collecting trace evidence.

Carina Hahn sits at a workstation and looks at a computer screen.

Spotlight: Measuring the Decisions of Human Facial Forensic Examiners

A white van with its sliding door open, parked in a lot, with mountains in the background. Decals on the side refer to scientific studies.

Researchers Analyze THC in Breath of Cannabis Smokers

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17 April 2024

Nanoparticle fix opens up tricky technique to forensic applications

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Peter J. Vikesland is in the Department of Civil and Environmental Engineering, Virginia Tech, Blacksburg, Virginia 24061, USA.

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Detecting trace quantities of substances in complex aqueous media, such as river water, sewage and treated drinking water, can be like looking for the proverbial needle in a haystack. Numerous sensitive methods have been developed 1 , but they often rely on complicated sample-processing and signal-acquisition steps, and are typically time consuming and costly. An approach known as surface-enhanced Raman spectroscopy (SERS) can detect single molecules 2 , 3 and has long been promoted as a promising alternative to such methods. But SERS is a notoriously finicky technique, with intrinsic variabilities, which makes it hard to use to quantify low concentrations of dissolved compounds. Writing in Nature , Bi et al. 4 report a digitized SERS method for detecting waterborne substances that addresses this fundamental issue.

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Article Contents

Molecular analysis of skeletal evidence, migrant identification, search, detection and recovery, commingling analysis, biomechanics of bone trauma, decomposition research, bone microscopy, isotope analysis, facial imaging, recent advances in forensic anthropology.

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Douglas H. Ubelaker, Recent Advances in Forensic Anthropology, Forensic Sciences Research , Volume 3, Issue 4, December 2018, Pages 275–277, https://doi.org/10.1080/20961790.2018.1466384

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Forensic anthropology involves diverse applications of anthropological knowledge to medico-legal problems. While the applications are evidence-driven, the available scientific methodology and foundation have developed through decades of research and experience. The roots of this field are anchored in comparative human anatomy but methodology has developed through experimentation, the assemblage of documented collections and databases and thoughtful research design. While forensic anthropology represents a mature scientific field, it continues to evolve and advance through new, innovative global research. Much of this progress is fuelled by issues encountered in casework. The unique evidence and problems presented in forensic cases call for the very best scientific approaches available. Usually, the correct approaches and solutions can be found in the existing scientific literature. However, sometimes the unique issues presented by the casework cannot be addressed adequately with the existing techniques. These situations stimulate forensic anthropologists to seek new solutions through targeted research.

This Special Issue presents research advances in several areas of forensic anthropology that have sustained rapid, recent progress. While our journals continually reveal new information in all aspects of forensic anthropology, several areas of investigation have registered particularly strong academic interest featuring innovative research.

Successful recovery and analysis of DNA has dramatically affected many areas of forensic science. In the field of forensic anthropology, molecular analysis can yield highly accurate information regarding the sex of the individual represented and provide positive identification [ 1 ]. Molecular approaches also can contribute to ancestry evaluation and species recognition. The use of DNA for positive identification has had a major impact on the practice of forensic anthropology and related fields of forensic science.

While the merits and contributions of DNA analysis are profound, many related issues express the need for new, innovative research and technological development. Frequently, evidence submitted for forensic anthropological analysis is not in pristine condition. In many cases, recovered remains are incomplete and/or extremely degraded due to criminal activity and/or taphonomic factors. Some site investigations produce only small fragments where even species is not apparent. Decisions need to be made regarding what areas of bone or tooth should be examined. Since DNA analysis is an expensive and destructive process, these decisions are critical and can affect the outcome of the case. Of course, decisions regarding the type of DNA analysis also are critical and largely driven by both the availability of the antemortem information and the nature of the evidence. Experimentation and casework experience have greatly improved approaches to these issues.

Deaths related to the global movement of undocumented people across national borders present major forensic challenges. Even within countries, identification of citizens can be difficult with incomplete evidence and lack of information regarding missing persons. These problems are greatly exacerbated when different countries are involved and the international movement of the person represented is not registered officially. Such cases call for extraordinary investigation, thoughtful forensic analysis and international communication. These efforts can strain the available local resources and often fall short of positive identification.

Recent years have witnessed remarkable efforts to address the identification of deceased, undocumented border crossers. These initiatives have involved international cooperation, careful exhumation procedures, comprehensive anthropological analysis and new techniques such as isotope analysis to identify the likely regions/countries of origin.

The entire process of forensic anthropological investigation begins with the procedures of search, detection and recovery. Improper or inadequate detection and recovery of human remains can compromise the downstream analysis and interpretation. While the traditional techniques of surface survey and excavation continue to be needed, new approaches, especially those using advanced technology offer significant advances.

Search procedures can be especially challenging when only very general information is available regarding the likely location of human remains. Topographic features can present limitations, especially with dense vegetation and other ground cover. Investigations of humanitarian and human rights issues can present special search and recovery challenges when information suggests that wells, cisterns, sewer systems, mass graves or disposal in water were involved. Confronted with these problems, researchers have devised innovative new approaches to improve the probability of success.

Secondary deposits of human remains or those that have sustained significant disturbance involve loss of normal bone articulation patterns. When multiple individuals are involved, the resulting commingling presents challenges to determine the number of persons represented and to assemble remains of individuals for analysis, identification and return to families. Traditional approaches to commingling problems have involved sorting by the type and side (left or right) of bone, age at death, bone size and maturation, sex and pathological conditions. In some skeletal assemblages, taphonomic indicators can be helpful as well.

Once obvious sorting has been completed, questions persist regarding bone morphology related to individuals. Could a robust femur relate to a robust humerus and represent one individual? Recent advances in commingling analysis address this issue. New databases and computerized techniques establish the probabilities that different bones could relate to the same individual. Applications refine the determination of the number of individuals represented and facilitate analysis aimed at identification.

A primary function of anthropological analysis relates to the interpretation of bone trauma. Anthropologists must differentiate the skeletal alterations representing perimortem trauma from those relating to antemortem injury, developmental features or postmortem and taphonomic factors. Assessment of the biomechanical factors involved plays a key role in any interpretation. Knowledge of biomechanical principles is required to explain fracture patterns and other alterations likely related to perimortem trauma. Interpretation of bone trauma can be challenging. Such challenges have led to greater understanding of the principles involved and experimental work designed to improve interpretation.

Major new initiatives in forensic anthropology have focused on decomposition research. Experiments involving both humans and non-human animals have revealed great detail about the process and variation of soft tissue decomposition and hard tissue alteration. In general, such research has elucidated the many factors that influence both the nature and timing of the decomposition process. Clearly temperature and location (surface, in-ground, aquatic, etc.) have long been regarded as key factors. Research has also indicated that soil conditions, moisture, body composition, body condition, presence of clothing or enclosures, funerary treatment and many other factors can influence the process. Such information is needed to properly assess time since death (post-mortem interval) and post-mortem events related to criminal activity.

In 1965, Ellis R. Kerley [ 2 ] published a technique that allowed age at death to be estimated from microscopic examination of features in human compact bone from the femur, tibia and fibula. Kerley's procedure involved the examination of primary osteons, secondary osteons, osteon fragments and the extent of remaining circumferential lamellar bone. This approach gained recognition due to its reported accuracy and the fundamental processes of bone formation and remodelling that it expressed. Since 1965, the technique has undergone many revisions and expansions for application to other bones of the skeleton. Research also has revealed how bone microscopic examination can provide useful information on many issues of forensic anthropological analysis.

For decades, analysis of elemental stable isotopes has offered key anthropological information related to diet. Stable carbon isotopes recovered from human tissues have revealed if diet focused on plants with a C 3 photosynthetic pathway or a C 4 pathway and the herbivores that fed upon them. Analysis of nitrogen isotopes provides insight into the trophic level of human diet. In anthropological studies of ancient populations, such information is crucial to interpretations of dietary and horticultural practices.

Recently, researchers have applied the concepts of isotopic analysis to examine the geographical origin of human remains. When unidentified human remains are recovered in forensic contexts, investigators question if they represent someone who lived in the area of recovery or from somewhere else. This question is especially relevant in cases involving terrorism and unidentified possible migrants. Using a battery of stable isotope analyses, researchers can determine if the isotopic signatures from the unknown match local baseline data. If not, attempts can be made to determine from what geographic area the unknown originated. This exciting new area of forensic science investigation depends on the assemblage of baseline data from appropriate geographic regions.

Forensic anthropologists relate to issues of facial imaging in facial approximation, craniofacial photographic superimposition and interpretations of surveillance images. Facial approximation refers to the process of estimating the living facial image of a person from the evidence presented by a recovered skull. This technique is used to reach out to the public for leads in missing persons that could culminate in identification using other methods.

Craniofacial photographic superimposition involves comparing a facial photograph of a missing person with a recovered skull. This technique is used primarily to exclude when photographs are available of a missing person thought perhaps to be represented by the recovered remains.

Recent research has focused on enhanced use of computers and related technology, as well as targeted efforts to clarify the relationship between soft and hard tissues. Facial approximation continues to represent a blend of art and science; however, recent advances have strengthened the scientific foundation.

Articles in this Special Issue of Forensic Sciences Research focus on overviews of the published literature on these topics. They also share results from the latest innovative research on these key areas of forensic anthropology applications.

Baker L . Biomolecular applications . In: Blau S Ubelaker DH , editors. Handbook of forensic anthropology and archaeology . 2nd. ed. New York: Routledge ; 2016 . p. 416 – 429 .

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Kerley ER . The microscopic determination of age in human bone . Am J Phys Anth . 1965 ; 23 : 149 – 163 .

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155 best forensic science research topics for your paper.

Forensic science or criminalistics applies scientific methodology and principles to solving crime and aid criminal justice procedures and laws. This area of study covers many fields ranging from computer forensics to doctoral research and forensic psychology.

For students specializing in forensic science studies, it is common to have to write an essay, research paper, or dissertation on the subject’s topics. The tricky part here is to select the perfect topic from a wide array of forensic science topics for a research paper. You could work on something that focuses on a neglected area of study in the field or go in for a controversial topic. You can also pick a common topic and throw new light on it, or simply choose a topic highlighting societal trends.

Whatever you choose to work on, it is essential to clearly state your research question/topic, offer defensible logic, have a well-elaborated body and a concise conclusion to score well.

Here is a list of some of the most interesting research topics in forensic science, which will allow you to write a good essay and score well. Take a look:

Forensic Science Research Paper Topics

These are some common but good forensic science topics that are sure to get you great reviews:

Hereditary in fingerprints and pattern similarities among family members
Is it possible to solve crimes with forensic dentistry?
Understanding the crime through the skeletal remains of the victim
Victim identification through skeletal remains
The association between legal ethics, forensic dentistry, and crime-solving
Techniques of gathering DNA sample through buccal cell collection in possibly violent subjects
Using forensic anthropology to identify relations, family, and ancestry
Using forensic anthropology to identify the age, gender, and size of the victim.
Forensic evidence – Types, chain of custody, collection, and analysis
Using forensic anthropology to identify trauma and disease history
Forensic ballistics and how the type of weapon used can be confirmed?
Are standard hair tests accurate in racial estimation and solving crimes
How to develop the best fingerprint testing powder for unmatched result clarity
The role of forensic psychology in assessing the probability and incidence rate of rape
Forensic toxicology and its role in analyzing the effects of anti-stress drugs on military members
The growing need for researching the technologies supporting forensic geomorphology
Blood spatter studies that aid forensic science in solving crimes
Forensic microscopy – an insight
Hair and fibers – What they say in forensic science crime cases
Poison chemistry – how forensic experts work
Forensic investigations of WW1 and WW-II war graves
Forensic reconstructions – importance, role, and scope in solving crimes
The laws and principles guiding forensic science
Sawn-off shotguns — evaluating pellet distribution pattern based on barrel length
Mitochondrial DNA Analysis of skeletal remains – An overview of the technique
Forensic ballistics — what a bullet may tell about the crime and the weapon used
Forensic evidence — assembling the pieces to the criminal jigsaw
Drug detection timelines — an insight
Development and validation of the method used to assess the quality of friction skin impression for evidence
Fatal intoxication through Isotonitazine – A case study

Forensic Science Research Topics For High School

These are excellent topics for high school students, which are easy to work on and create impressive essays related to forensic science:

Comparing sibling fingerprints with strangers
Finding out if fingerprints of identical twins are also similar
Differences to quickly separate human skeletal remains from those of animals
Victim identification – What you must know about facial skeleton
DNA testing – uses and limitations
Different types of stab injuries are important from a forensic science perspective
The dental pattern of wild animals and ways to recognize them.
Blood spatter patterns and what they indicate about the crime
Creating blood spatter lab for better understanding of spatter patterns
Forensic science autopsy reports and what they indicate
Careers in forensic science and upcoming specializations
Forensic science and analysis of fiber evidence from the crime scene.
Good practices in forensic science laboratories
Handwriting analysis and how it helps forensic science professionals
The role of Forensic Genealogy in solving age-old cases
Iodine fuming to reveal latent fingerprints
Immunoassay techniques for protein identification
Thin Layer & Paper Chromatography to identify materials
Forensic art as a means to identify suspects
Gas chromatography to identify liquids

Interesting topics To Research That Have to Do With Forensic Science

These research topics related to forensic science will help you create an exciting write-up that will draw attention to your knowledge in the subject:

Studying the composition of cling film used to package illegal drugs to separate drug traffickers from consumers
Dense materials and their role in Geoforensics
DNA typing – Pros and cons from a crime-solving perspective
Using gene expression to assess the age of injury
Differences in stab injuries that confirm if wounds were antemortem or postmortem
The study of glass fracture pattern to assess bullet firing direction and distances
Ground-penetrating radar systems and their role in Geoforensics
Gamma-ray radiography and its role in crime-solving geomorphology studies
What insects tell us about the murder scene
The correct way to prepare a forensic autopsy report
Forensic psychology and law – an insight
The forensic science behind fire investigations
Organic chemistry and its role in forensic science for solving crimes
Mathematics and statistics — how they aid cases of forensic science
The forensic science behind identifying forgery and counterfeiting
Use of 3D imaging for visualization of footwear and tire impressions at the crime scene
Fluid dynamics study to assess bloody fingerprints at the crime scene
Audio recordings — interpretation and processing to build a case
Recorded gunshot sounds and how they are interpreted and analyzed to solve crimes
Cartridge case comparisons to assess pressure factors in firearms

Controversial Topics In Forensic Science

This is a collection of research paper topics for forensic science that is sure to spark a debate when discussed in class:

Are changes in the composition of cling film used for drug packaging of any forensic value to arrest criminals?
DNA typing — an analysis of the efficacy of this technique in identifying victims and unsolved crimes
Isotope ratio mass spectrometry — an analysis of accuracy concerning the crime, its conditions, and people involved
Flies that come on a dead body based on its location and time of death
Forensic psychology and its impact on instances of Military rape
Is forensic toxicology research lacking in assessing drug abuse among military personnel during combat?
Is Forensic geomorphology helpful in solving crimes?
Skeletal tissues and forensic microscopy — the role they play in solving crimes
Forensic science and how it makes the dead speak
Characteristics of Zopiclone degradation and consequences from a forensic toxicology perspective
Understanding what happens to the body when someone dies naturally vs. a violent death.
Criminal minds and their forensic psychology
The role of Trace DNA in criminal investigations and its reliability
Criminal profiling and the role of Forensic psychology in arresting the killer
Forensic victimology and its role in context with investigations and legal parameters

Forensic Science Topics For Presentation

Suppose you want to make a presentation on any aspect of forensic science. In that case, you can consider the topics given below as they provide sufficient scope and information:

How to extract fingerprints from a crime scene
Equipment used in crime scene fingerprinting and the process
How to identify skeletal remains of a human subject
Building the face of a human through the remains of their skeleton
Know all about the various techniques used in forensic anthropology
Cyber forensics – How to catch a hacker
Methods of DNA testing
Methods of DNA testing of unwilling subjects
Standard forensic hair tests used in forensic examinations
Identification of cell type and body fluid through RNA based methods
Types of flies that congregate on a dead body indicating the time of death
A detailed step-by-step guide to forensic deduction
Cyber Forensics – how to detect phishing
Forensic science – the basics of crime scene analysis
Forensic frauds and laws around examiner misconduct
Methods and tools used by forensic pathologists
Forensic autopsy Vs. Regular Autopsy
The Muscid fly and its forensic importance
The role of forensic science in tracking victims of human trafficking
Evaluation of footwear impression — a step by step guide of the Footwear Impression Comparison System (FICS)

Forensic Science Thesis Topics

Use any of the topics given below to write an impressive thesis that showcases in-depth knowledge. These topics provide ample scope to delve deeper into the subject and write after thorough research.

Fingerprint science — an insight
Crime scene fingerprinting — a detailed study
Forensic anthropology — an insight
Forensic anthropological techniques for a detailed history of the victim
Isotope ratio mass spectrometry — An Insight
Computer forensics and its role in solving cybercrime
Forensic toxicology and how it may help prevent military violence
The role of forensic anthropology in postmortem findings
DNA testing methodology — an insight into means available and their efficacy
Reassessment of Asphyxia and the Pink Teeth Phenomenon
Forensic psychology – differentiating sociopaths from psychopaths
Study of blood in forensic science
DNA typing – An insight into unsolved crimes and the use of this method
Nanotechnology in Forensic studies – An insight
Consumer protection in the world of growing cyber crimes and how cyber forensics can help
Forensic toxicology – An insight
Fingerprint ridge density and its relation with sex determination
Forensic geomorphology – an extensive study of the scope and application of this field of study
The history and evolution of forensic science
Medical Image security using digital watermarking – An overview
Anatomy of bruises and what they convey about the tracker and the victim
Forensic psychology – Understanding the characteristics of psychopaths for profiling.
High-Resolution Melt (HRM) assays – An insight
Human vaginal matrix and the analysis of condom evidence from a forensic perspective
Handprint dimensions for Sex classifications – the ABC model of forensic science

Current Topics in Forensic Science

These topics stem from the latest developments in forensic sciences and highlight the current environment in this field of study. Take a look:

Latest techniques in forensic science to catch murderers
Studying injury age using gene expression to solve the crime
Latest forensic science techniques used to identify illicit drugs
Developments in technology and chemistry that aid forensic science like never before
Latest illegal drug recognition systems – an insight
The amino acid fingerprint test – an evaluation
Identifying of Felidae Animals using Elemental Analysis of Hair and its Scanning Electron Microscopic Characterization
Pediatric Poisoning — Seasonal trends, materials and distribution across the UK.
The UK forensic science environment and what has changed
The changing scene of Geomorphology with gamma-ray radiography and ground-penetrating radar
How stable is the use of Zopiclone in forensic studies related to whole blood samples
Latest enhancements in RNA based methods for identification of cell type and body fluid
The emerging role of computer forensics in cybercrime and the latest developments.
Cyberwarfare and how cyber forensics fit in?
Geoforensics – how soil mapping aids in solving crimes
mRNA profiling assays and their role in reducing time and cost of analysis
Latest technologies used in the identification of body fluids
Latest trends in forensic odontology
Heartbreak Grass deaths — A toxicological and medico-legal analysis
Water-soluble paper analysis for forensic discrimination
A worldwide survey of issues in forensic science – A contemporary perspective
DNA study to assess regular occupancy in any vehicle
Methods of assessing authentic audio recordings using different voice apps
An insight into the DNA extraction process for DNA identification in bodies
One-handed strangulation – A reconstruction study to find the criminal

If you are looking to submit an impressive dissertation or essay on a topic on forensic science and are still confused about how to proceed, get in touch with us. We can help you find impressive topics for your dissertation and help with forensic science research proposal topics. We have a strong team of expert writers and provide an array of high-quality, professional writing services for college and high school students.

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Blood test can detect laughing gas and help the police fight whippit abuse

by University of Copenhagen

Forensic blood analysis can reveal whether drivers are high on laughing gas. Researchers hope the method can help the police.

Once again, recreational use of laughing gas has attracted media attention in Denmark. This time, however, focus is on drivers who have inhaled laughing gas before getting behind the wheel—with potentially lethal consequences.

So says Ph.D. Student of Forensic Chemistry Angelica Ørregaard Lindholm at the University of Copenhagen, who is responsible for the new study published in Forensic Science International on driving under the influence of laughing gas.

"Laughing gas can cause disorientation, hallucinations, dizziness, and fainting. So, laughing gas and driving is a dangerous combination," Ørregaard Lindholm explains and adds that it is a common misconception that the drug does not show up on tests.

In the new study, Ørregaard Lindholm and her colleagues show that laughing gas can be detected in a simple blood test .

"We have analyzed 62 blood tests, 52 of which were positive for laughing gas. The police have provided the tests, which are made when they have reason to suspect someone of being under the influence of laughing gas," Ørregaard Lindholm says.

After publishing the study, the researchers have continued analyzing tests. Of the 103 tests they have analyzed since 2020, 90 have been positive.

Since 1 July 2023, it has been illegal, with a few exceptions, to carry laughing gas in public spaces , and this includes cars in public spaces. Being high on laughing gas is not illegal, though.

Laughing gas is also known as nitrous oxide and can be bought in small or large cartridges intended for whipped cream siphons.

"Inhalation of laughing gas can induce short-term euphoria and a sense of weightlessness," Ørregaard Lindholm says.

It is used as a painkiller by dentists and during labor, and here it is safe to use. However, long-term abuse can cause permanent neurological damage.

"Nitrous oxide inactivates the vitamin B12 stores in the body, and they take time to rebuild. If you continue to use laughing gas, your body will begin to suffer from vitamin B12 deficiency, and this will damage the protective myelin sheath that surrounds the nerve fibers," Ørregaard Lindholm explains.

Method must be refined

Even though laughing gas can be detected in the blood, the researchers face several challenges before they are satisfied with the method.

"Our main problem is that the effect is brief and that the gas disappears quickly after inhalation," Ørregaard Lindholm says.

Therefore, the police must act quickly. Otherwise, the gas will not show up on the test.

"This is likely to limit the police's options. For instance, if they are in the countryside and the nearest hospital is far away, they might think that testing is futile and decide not to do the test," Ørregaard Lindholm explains.

Research from the 1970s shows that the half-life of laughing gas is five minutes. Sometimes, though, the researchers found nitrous oxide in the blood three hours after the sample was taken.

Now they want more systematic knowledge of how long laughing gas can be detected in the blood and to which extent it affects the user's ability to drive.

Therefore, the next step for Ørregaard Lindholm and her colleagues is to do a clinical trial with a group of test subjects who will be given laughing gas and asked to operate a car simulator while the researchers do a series of blood tests. The trial will be conducted together with the Center of Head and Orthopaedics at Rigshospitalet and the Division of Transport Science at the Technical University of Denmark.

"Ultimately, we hope our method and the results of the clinical trial will be able to help the police get the proof they need in cases of laughing gas abuse and driving."

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April 22, 2024

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Advance in forensic fingerprint research provides new hope for cold cases

by Meg Cox, Loughborough University

New hope for cold cases due to breakthrough in forensic fingerprint research

Researchers have unveiled a method capable of detecting drug substances from fingerprints lifted from crime scenes, which could provide fresh insights into unsolved cases. The research is published in the journal Drug Testing and Analysis .

Analytical scientists from Loughborough University have demonstrated for the first time that drug residue—namely the fast-acting sleeping pill Zolpidem, which has been linked to drug-facilitated sexual assault and drink spiking —can be detected on gel-lifted fingerprints.

Dr. Jim Reynolds and Dr. Ayoung Kim say the breakthrough could shed new light on cold cases and unsolved crimes as forensic gel lifters—which transfer prints onto a gelatin surface—are used globally by scenes of crimes officers to preserve and visualize fingerprints.

"This is the first time that analysis of gel-lifted prints for a drug substance has been accomplished, and shows that lifted prints and other forensic marks can be interrogated for useful information," says Dr. Reynolds, the research lead.

"Since gel-lifted prints and marks can be stored for many years, the technique could be of real use in cold cases where additional information may prove useful to either link or exonerate a suspect to the investigation. Working with police forces and applying the method to cold case samples could help bring criminals to justice who may have thought they have got away with it."

A number of tests exist to detect drugs directly from fingerprints, but these face limitations. They can be destructive to the fingerprint, degrade drug residues, and be affected by environmental interferences.

It has long been speculated that gel-lifted prints contain valuable chemical information and could offer more accurate drug detection.

However, traditional techniques used to analyze the chemicals present in a sample have previously not been suitable for gel lifters. This is because they detect all chemicals present, including those that make up the gel, making it difficult to identify specific substances.

The method used by Dr. Reynolds and Dr. Kim, called sfPESI-MS, overcomes this issue using a rapid separation mechanism that distinguishes the drug substance from the background of the gel.

This separation method enables the direct detection of a drug substance using mass spectrometry , a technique that identifies chemicals by measuring their molecular weight. The researchers have successfully tested the technique using Zolpidem-laced fingerprints lifted from glass, metal, and paper surfaces in a laboratory setting.

They now hope to work with police forces to analyze stored gel-lifted prints and use the method to identify other substances.

Dr. Reynolds said, "Zolpidem was the focus of our research, but the method could just as easily be applied to other drug substances a person may have been handling and could be applied to other chemicals such as explosives, gunshot residues, paints, and dyes.

"By linking chemical information to the fingerprint, we can identify the individual and link to the handling of an illicit substance which may prove useful in a prosecution. This could be useful to detect individuals who have been spiking drinks; for example, if the drug they are using gets onto their fingertips, then they will leave evidence at the scene."

Dr. Kim, who is the first author of the paper and completed the research as part of her Ph.D. at Loughborough, added, "We would like to apply our method to real samples from criminal investigations; it would be good to know my Ph.D. research has helped bring criminals to justice."

Journal information: Drug Testing and Analysis

Provided by Loughborough University

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Major milestone reached for key weapons component

Sandia National Laboratories and the Kansas City National Security Campus completed a crucial weapons component development milestone, prior to full rate production.

The Mark 21 Replacement Fuze interfaces with the W87-0 warhead for deployment onto the Minuteman III and, eventually, the Sentinel Intercontinental Ballistic Missile.

The first production unit of the replacement fuze was approved through the National Nuclear Security Administration's rigorous Quality Assurance Inspection Procedure in March.

The Mark 21 fuze, an integrated assembly of Sandia-designed logic and sensing devices, directs the warhead to initiate firing sequences at the correct point in targeting parameters. The Mark 21 Fuze Replacement Program is a Strategic Partnership Program effort between the NNSA and the U.S. Air Force.

"Sandia's role is to deliver a fuze that meets the Air Force's requirements. Completing the first production unit is a visible milestone in maintaining a credible deterrent," said Brad Boswell, a director in Sandia's nuclear deterrence modernization program. "It demonstrates that we are providing the necessary capability for the U.S. nuclear deterrent into the future."

Designing the fuze

As the lead design agency of the Mark 21 fuze, Sandia was responsible for incorporating Air Force requirements into a form, fit and functional package that interfaces with the Mk21 aeroshell, MMIII missile and W87-0 warhead. Work began in 2011.

"The early engineering work takes a lot of time. While some of it is done in parallel, much of the work must be sequential," said Bob Oetken, a former program manager for the fuze program. "We're doing a very specific job that requires, in many cases, custom parts."

Sandia worked closely with the Kansas City National Security Campus to ensure the design was producible and delivered on schedule. While the Kansas City National Security Campus is producing and assembling the fuze, Sandia is manufacturing some of the parts.

Fuze qualification

Before production of the fuze began, a significant amount of time was spent qualifying the design, which ensures the design meets Air Force specifications. It includes analysis through testing and other engineering tools to ensure the fuze will function as intended.

"Qualification involves significant resources and time. We are in a very rigorous business. Some of the qualification is used to verify that we are meeting requirements and took as long as three years from concept to delivery," Oetken said.

The program has conducted numerous tests, including environmental, ground and flight tests, to ensure the fuze withstands multiple difficult environments, such as acceleration, vibration and thermal environments expected during missile launch and ballistic reentry.

Stockpile assessment

Now that the fuze is in full rate production, Sandia's responsibilities shift.

"As the design agency, Sandia is responsible for the component until the weapon is retired. Sandia is accountable for ensuring that the weapon with the fuze attached is safe, secure and reliable," Boswell said.

As part of the stockpile assessment, Sandia will conduct surveillance through processes such as modeling and simulation and flight and ground testing to ensure the fuze continues to work as intended.

Partnering for great results

Completing the first production unit of the Mark 21 fuze is the culmination of more than a decade of work and collaboration, not just at Sandia, but by NNSA and the Department of Defense.

"This first production unit marks a significant milestone not just for Sandia but the broader nuclear security enterprise, particularly our partners at the Kansas City National Security Campus," Boswell said. "While we stop to recognize this achievement, we must also acknowledge this is the start of the next phase of our partnership as we work to meet key production milestones to deliver the quantity of fuzes needed to meet the needs of the stockpile."

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