Forensic Science International-Genetics最新文献

{"title":"Development and evaluation of an optimized Ancient DNA extraction method for femoral diaphyses and simulated heat-treated teeth","authors":"Guihong Liu ,&nbsp;Shengqiu Qu ,&nbsp;Qiushuo Wu ,&nbsp;Yazi Zheng ,&nbsp;Mengyu Tan ,&nbsp;Jiaming Xue ,&nbsp;Haoyan Jiang ,&nbsp;Mengna Wu ,&nbsp;Yuxuan Tan ,&nbsp;Miao Liao ,&nbsp;Jing Yan ,&nbsp;Meili Lv ,&nbsp;Lin Zhang ,&nbsp;Weibo Liang","doi":"10.1016/j.fsigen.2025.103371","DOIUrl":"10.1016/j.fsigen.2025.103371","url":null,"abstract":"<div><div>Extracting endogenous DNA from highly degraded hard tissue samples, such as bones and teeth, is crucial yet challenging for forensic human identification. Existing forensic methods and commercial kits for DNA extraction from bones and teeth often inefficient for old, poorly preserved, chemically treated, or heat-exposed samples. Ancient DNA (aDNA) extraction technology is highly effective in recovering degraded, short DNA fragments and has been successfully used to isolate DNA from skeletal remains and sediments dating back to tens of thousands of years. However, its direct application to individual identification and short tandem repeat (STR) analysis may require further adaptation to meet specific forensic requirements. Therefore, this study was conducted to optimize aDNA extraction techniques for developing a method tailored for highly degraded forensic samples. We developed a forensic aDNA-based extraction (FADE) method by refining the aDNA extraction protocol through optimization of lysis and purification conditions. The effectiveness of the developed method was validated using 8 forensic case samples (femoral diaphyses) and 20 simulated heat-treated teeth. For samples subjected to prolonged heat treatment, the FADE method significantly increased peak heights, with improvements of 30 % and 45 % in the 30- and 40-min heat-treatment groups, respectively, and yielded a greater number of amplified loci and alleles. The FADE method developed in this study is well-suited for forensic applications involving highly degraded femoral diaphyses and simulated heat-treated teeth, significantly enhancing the success rates of DNA analysis and providing an effective technical solution for complex forensic DNA extraction.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103371"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145245551","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Establishment and application of a triplex RTqPCR assay system for the identification of forensic body fluids","authors":"Shuxiao Hu ,&nbsp;Liping Chen ,&nbsp;Jiayao Duan ,&nbsp;Sheng Hu ,&nbsp;Yixia Zhao ,&nbsp;Yang Li ,&nbsp;Ruiqin Yang ,&nbsp;Anquan Ji ,&nbsp;Jie Lian ,&nbsp;Qifan Sun","doi":"10.1016/j.fsigen.2025.103337","DOIUrl":"10.1016/j.fsigen.2025.103337","url":null,"abstract":"<div><div>MicroRNAs (miRNAs) have emerged as valuable biomarkers for the identification of forensic body fluids due to their stability and tissue specificity. However, the limited quantity of body fluid at crime scenes often hampers the accuracy of miRNA-based detection methods. In this study, we developed a triplex reverse transcription-quantitative polymerase chain reaction (RT-qPCR) assay system that enables the simultaneous detection of three miRNAs, improving throughput and efficiency while overcoming challenges in forensic investigations. First, the primers and probes of the miRNAs were redesigned to meet multiple detection requirements on the basis of a previous study in which five types of body fluid-specific miRNAs and internal genes (miR-451a, miR-891a-5p, miR-144–5p, miR-203a-3p, miR-223–3p and miR-320a-3p) were screened in a laboratory. The primer and probe concentrations, premix concentration and annealing temperature were subsequently optimized to establish a triplex RT<img>qPCR assay system. This system enables the simultaneous reverse transcription of six miRNAs from a single sample, followed by two separate triplex amplification reactions to quantitatively analyze all six miRNA markers. The amplification efficiency, primer cross-reactivity, repeatability, triplex detection and single detection results of the system were subsequently analysed, and a prediction model was constructed by combining the sample data with a kernel density estimation (KDE) method. Finally, the ability of the detection method to identify body fluids was further verified with authentic samples. The results demonstrate that the optimized triplex RT<img>qPCR assay system achieves the same detection performance as the single detection system, but is faster and more cost-effective. This technology is especially suitable for the detection of trace body fluid stains left at crime scenes and effectively solves the contradiction between the requirements of repeated RT<img>qPCR detection of traces and multiple sample sizes. In addition, the body fluid identification model, which was established by the data obtained from the triplex RT<img>qPCR system combined with KDE, was successfully applied to predict and identify simulated samples and actual samples. This system provides an effective tool for the identification of suspicious body fluids and lays the foundation for further research on multiplex RT<img>qPCR assay systems and the construction of more accurate data models.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103337"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144864760","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparison of DNA profiles from samples collected from underneath fingernails and hand deposits following everyday activity","authors":"Mariya Goray ,&nbsp;Adrian Linacre ,&nbsp;Roland A.H. van Oorschot ,&nbsp;Duncan Taylor ,&nbsp;Kahli Murton","doi":"10.1016/j.fsigen.2025.103367","DOIUrl":"10.1016/j.fsigen.2025.103367","url":null,"abstract":"<div><div>In instances of direct physical contact between individuals involved in criminal activity, body samples can provide significant and relevant information to aid in criminal investigations and court proceedings. Fingernails are one such forensically relevant body area that is capable of providing evidence of direct contact and potentially revealing whether the interaction involved was of a forceful kind. Several studies have investigated the prevalence of non-self DNA under fingernails after different crime-related scenarios; however, few have assessed the types of DNA profiles found after everyday activities. Furthermore, the comparability of the fingernail samples to those deposited on contacted surfaces remains unknown. In this study, we examined the composition of self- and non-self-DNA in samples collected from under the fingernails and held tubes from the same set of individuals. Additionally, the potential use of fingernails samples for shedder assessment was evaluated through comparison with two common shedder categorisation tests. For these purposes, samples were collected from both hands of 25 individuals of different demographics, without any prior restrictions on activities. Direct deposits were made by holding a 50 mL tube (for DNA shedder testing) and placing index fingers onto a slide (for Diamond™ dye cell counting shedder testing). Fingernail samples from both hands were taken immediately after tube-holding deposits. Reference DNA samples were collected from the participants as well as their cohabitating partners and other adults. Qualitative and quantitative data on DNA and cell deposits were collected to support activity-level evaluations. In our study, mixture inversions were rare, with non-self DNA, when detected, usually present as a minor component. More non-self DNA was detected after participants’ contact with the tube compared to fingernail samples. Partners’ DNA was frequently detected in both sample types, but more so in fingernail samples. Comparisons of the three shedder testing methods (fingernails, tube holding and cell count) showed that the categorisation results of these methods are not interchangeable and that DNA methods (tube vs. fingernails) were more consistent (64 % of deposited classified into the same shedder category) with each other than with cell counts (tube vs. cell count: 52 % classified into the same shedder category) (fingernails vs. cell count: 40 % remained in the same category). We anticipate that these datasets will serve as a valuable resource for activity-level evaluations and encourage other investigators to contribute to the growing data collection.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103367"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145202513","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pharmabiome analyses in tandem with chemometrics can help trace the provenance of falsified medicines: A proof-of-concept study","authors":"Carla Perez-Mon ,&nbsp;Alberto Roncone ,&nbsp;Aiman Abrahim ,&nbsp;Marivil Islam ,&nbsp;Cathrin Hauk ,&nbsp;Celine Caillet ,&nbsp;Hamid A. Merchant ,&nbsp;Rabia Farzand ,&nbsp;Luana Bontempo ,&nbsp;Simon D. Kelly ,&nbsp;Daniel Blessborn ,&nbsp;Joel Tarning ,&nbsp;Rachel Kline ,&nbsp;Victoria Nicheva ,&nbsp;Dominic T. Kurian ,&nbsp;Paul N. Newton ,&nbsp;Rob Ogden","doi":"10.1016/j.fsigen.2025.103392","DOIUrl":"10.1016/j.fsigen.2025.103392","url":null,"abstract":"<div><div>A lack of robust analytical approaches limits our ability to investigate the clandestine manufacturing origins of falsified medicines. We conducted a proof-of-concept study to test the feasibility of geolocating the production sites of falsified medicines, based on the identification of site-specific biological and chemo-isotopic features using a combination of environmental DNA metabarcoding, Direct Analysis in Real Time - Mass Spectrometry and Isotope Ratio Mass Spectrometry as profiling techniques. We produced tablets at two distant locations (England vs. Thailand), using controlled manufacturing methods, excipient composition and environmental conditions. Sets of tablets produced at separate locations showed distinct bacterial and eukaryotic diversity, particularly influenced by the incorporation of water used during tableting and the background environmental biosignatures of the production site. Tablets showed corresponding site-specific chemometric profiles, but the factors contributing to the observed chemical differences were unclear. When reference samples of known origin are available, our study suggests that site-specific biological and chemical features can be used in modelling approaches to successfully predict product origin. We developed a new mapping approach to exploit the geographic information within the eukaryotic pharmabiome of the falsifications; based on eDNA-derived species identification and the integration of publicly available species distribution data. In the absence of reference samples of known origin, the application of this workflow to our dataset provided partial clues about the product’s origin, with limitations likely due to taxonomic resolution and the presence of species with wide distribution ranges. Collectively, our research provides experimental support for the development of integrated, multifaceted tools for tracing the origin of falsified medicines, advancing efforts to combat this pervasive but neglected global health problem.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103392"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145571583","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Epigenetic differentiation of monozygotic twins in forensic investigations using nanopore sequencing","authors":"Kuo Zeng ,&nbsp;Kai-bo Yang ,&nbsp;Jiang Du ,&nbsp;Allah Rakha ,&nbsp;Zhenze Liu ,&nbsp;Dan-yang Wang ,&nbsp;Yun-zhou Chen ,&nbsp;Si-wen Wang ,&nbsp;Mao-ling Sun ,&nbsp;Hongbo Wang ,&nbsp;Yi-long Wang ,&nbsp;Atif Adnan ,&nbsp;Jun Yao","doi":"10.1016/j.fsigen.2025.103344","DOIUrl":"10.1016/j.fsigen.2025.103344","url":null,"abstract":"<div><div>Monozygotic twins (MZTs) pose a significant challenge in forensic genetics due to their identical nuclear DNA, rendering conventional markers like STRs and SNPs ineffective. This limitation highlights the need for novel approaches to distinguish MZTs in criminal investigations, disaster victim identification, and paternity testing. Current methods lack the resolution to differentiate MZTs, creating a gap in human identification. Epigenetic markers, such as DNA methylation, offer a solution by reflecting environmental and stochastic differences post-twinning. However, traditional sequencing technologies often fail to meet forensic requirements for resolution, speed, or reliability. To address this, we employed Oxford Nanopore sequencing to analyze genome-wide DNA methylation patterns in six MZT pairs, identifying robust epigenetic biomarkers for forensic discrimination. Our approach leverages long-read sequencing and single-base resolution to overcome conventional limitations. The study aimed to identify differentially methylated loci (DMLs) as stable, heritable biomarkers for distinguishing MZTs, even in degraded or trace samples, while demonstrating the forensic utility of nanopore sequencing. We identified 3820 shared DMLs enriched in metabolic and neural pathways, localized to promoter (1.84 %) and intergenic (88.03 %) regions. Nanopore sequencing achieved &gt; 99.5 % alignment efficiency and &gt; 13 kb N50 read lengths, enabling rapid, PCR-free analysis. This study demonstrates that nanopore-based methylation profiling effectively distinguishes MZTs by capturing environmentally influenced epigenetic differences, providing actionable biomarkers for forensic discrimination. By bridging epigenetics and forensics, our findings advance precision in human identification, offering transformative tools for criminal casework, disaster victim identification, and paternity testing, addressing a longstanding limitation in resolving cases involving genetically indistinguishable individuals.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103344"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144888946","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"DNA methylation-based forensic framework for age prediction and body fluid identification using nanopore sequencing","authors":"Isis Beentjes ,&nbsp;Martin A. Haagmans ,&nbsp;Desiree D.S.H. de Bruin ,&nbsp;Anwar Permana ,&nbsp;Ewelina Pośpiech ,&nbsp;Wojciech Branicki ,&nbsp;Amade Aouatef M’charek ,&nbsp;Kristiaan J. van der Gaag ,&nbsp;Titia Sijen ,&nbsp;Peter Henneman","doi":"10.1016/j.fsigen.2025.103370","DOIUrl":"10.1016/j.fsigen.2025.103370","url":null,"abstract":"<div><div>Forensic DNA phenotyping applies genetic and epigenetic markers to infer biogeographical ancestry, physical appearance traits, and biological age, particularly when conventional methods fail to identify a suspect or victim. DNA methylation, a key epigenetic modification, is especially valuable for age estimation and due to its tissue-specific patterns also for body fluid identification, with the latter aiding in crime scene reconstruction. Current MPS-based methylation sample analyses require time-consuming, multi-step processing, while Nanopore sequencing offers a promising alternative by enabling real-time, direct detection of methylation without DNA conversion. This study evaluates the potential of Nanopore’s PromethION 2 platform for comprehensive, single-assay forensic epigenetic analysis of biological age estimators (epigenetic clocks) and body fluid markers, focusing on relatively low DNA samples (&lt;100 ng.) Our results showed that low read depth coverage can invoke the occurrence of beta values equal to zero and one for the methylation status (whilst methylation has a continuous nature), provide challenges for accurate age estimation and body fluid identification. Our study demonstrates that both the age prediction models tended to estimated older ages than expected; however, applying a proof-of-concept linear correction model significantly enhanced age estimation accuracy. Body fluid identification of blood and saliva was highly accurate in conditions with high and low read depth coverage, correctly identifying 4/4 sample in each experiment. This explorative study highlights the potential of adaptive sampling on PromethION for forensic age prediction and body fluid identification, while future studies should focus on validating analysis thresholds, improving lower-quantity sample performance, advancing body fluid identification models for an extended tissue set, and mixture analysis.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103370"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145294908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Preservation of proteomic and genomic genotypes in skeletal material from forensically treated cadavers","authors":"Glendon Parker ,&nbsp;Michelle Harrel ,&nbsp;Jennifer Snedeker ,&nbsp;Michelle Salemi ,&nbsp;Trevor Borja ,&nbsp;Rachel Houston","doi":"10.1016/j.fsigen.2025.103384","DOIUrl":"10.1016/j.fsigen.2025.103384","url":null,"abstract":"<div><div>Skeletal material is often the only remaining evidence in missing persons cases, for mass burials, or disaster victim identification. Nuclear DNA can be highly degraded in these samples making it difficult to recover quality profiles for human identification or to deconvolute comingled remains. Proteins are chemically more stable than DNA and contain genetic information in the form of genetically variant peptides (GVPs) that can be used to infer non-synonymous SNP genotypes. To test chemical and genetic preservation of both biomolecular types, eight cadavers were examined using five treatments: embalming, burning, burial, and surface decomposition with and without subsequent formalin treatment (0–66 h in 36 % (w/v) formalin). Across the study DNA recoveries ranged from 210 ng to 0.08 ng in 50 mg of processed bone, a 2,600-fold range. Peptide recoveries, ranged from 22,000 ng to 5900 ng from the same amount of bone, a 3.7-fold range. At the information level the same peptide input (750 ng) could be obtained for all samples, and nucleotide level (0.8 ng) for most samples, although this consumed more sample than proteomic analysis. Of the 18 STR-profiles analyzed, 9 were CODIS-ineligible, yet all samples contained GVPs. This indicated that GVPs can be used to construct non-synonymous SNP genotypes in degraded bone samples that cannot provide usable STR-profiles. The relative quality of proteomic and genomic genetic data had a linear relationship (P &lt; 0.0001) with a loss of 1 GVP for a 14 % loss of STR-alleles (97 % CI = 10–22 %), indicating that even correcting for chemical loss, information quality still decreased. SNaPshot™ genotyping of non-synonymous SNPs was less sensitive than GVP inference. Proteomic analysis of individual proteins in the sample indicated that collagen degradation was responsible for most protein loss, and chemical modifications were modest. Future studies are required to examine the stability of both DNA and protein in specific cadaver treatments, for further GVP discovery and validation, and optimization of proteomic sample processing. However, given the stability of GVP detection, even at this preliminary stage, protein-based genotyping is a valid option for extracting genetic information from samples with minimal or even no usable DNA detected. This is the first example of successful proteomic genotyping in forensically degraded bone and demonstrates that proteomic genotyping can be used for human identification and potential deconvolution of comingled remains.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103384"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145472502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Human DNA recovery in the context of wildlife crime: Comparison of trace DNA collection methods from wildlife specimens","authors":"A. Thomas ,&nbsp;S. McColl ,&nbsp;Robbie Rae ,&nbsp;R. Ogden ,&nbsp;L. Gibson ,&nbsp;N. Dawnay","doi":"10.1016/j.fsigen.2025.103361","DOIUrl":"10.1016/j.fsigen.2025.103361","url":null,"abstract":"<div><div>Forensic DNA analysis, used for the purposes of species, sex, individual, and, geographic determination of wildlife is one of the most applied forensic techniques in, wildlife crime investigations. However, in most other criminal investigations forensic, DNA analysis refers to human DNA profiling for the purposes of identifying victims, and/or perpetrators. The ability to recover human DNA profiles from the surfaces of, wildlife specimens, such as ivory or fur, opens up opportunities for identification of, individuals involved in wildlife crimes in the absence of other evidence types. This, study aimed to compare the effectiveness of four different human touch DNA recovery, methods, cotton swabs, flocked swabs, foam swabs, and minitapes, from the surfaces, of a range of wildlife derivatives. Groups of four participants handled ivory, elephant, skin, snake skin, conch shell, antler, and antelope fur. DNA was subsequently, collected extracted, quantified, and profiled. Foam swabs, a non-traditional method of, touch DNA recovery, recovered the highest average DNA concentrations and number, of alleles across all specimen types acting as an effective cross-substrate recovery, method. Flocked swabs performed as the second-best recovery method for all, specimens except when sampling from antelope fur. Minitapes and cotton swabs, showed comparatively poor performance during this study despite being the two most,common DNA recovery techniques currently employed by law enforcement. Ivory, yielded the highest average human DNA concentrations but paradoxically produced a, significantly lower number of donor alleles. Our results indicate fresh touch DNA, deposits are recoverable from multiple wildlife specimens and recommend that, attempted recovery of touch DNA should be a routine consideration by forensic, practitioners during wildlife crime investigations.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103361"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145049078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microhaplotypes in forensic genetics: From exploration to application in degraded DNA specimens","authors":"Chiara Turchi ,&nbsp;Filomena Melchionda ,&nbsp;Fabiano Gentile ,&nbsp;Alberto Marino ,&nbsp;Domenico Colloca ,&nbsp;Mauro Pesaresi ,&nbsp;Andrew J. Pakstis ,&nbsp;Kenneth K. Kidd","doi":"10.1016/j.fsigen.2025.103391","DOIUrl":"10.1016/j.fsigen.2025.103391","url":null,"abstract":"<div><div>Microhaplotypes have emerged as powerful forensic markers over the past decade. This paper sets out the development of a MPS panel of microhaps and its potential for application to identification, analysis of degraded DNA, ancestry inference, and identification of close biological relationships. To make it more effective when dealing with fragmented DNA, the MPS assay was designed to ensure a reduced amplicon size of less than 140 bp. After MPS assay validation, a panel of 76 microhaps, comprised of 299 different SNPs and spread across the autosomal human genome, was established. A total of 102 Italian individuals were analyzed to estimate the genotype and haplotype frequencies. The effective number of alleles at each locus (A_e) for the Italian population ranges from 1.926 to 6.187, with 59 MHs that have values greater than 3.0. The matching probability (PI) ranges from 0.055 to 0.345 and the cumulative PI value is 11.763E-66. Complete and reliable profiles were obtained with as little as 0.05 ng. The MHs panel was then validated on real forensic specimens chosen on the basis of their DNA content and degradation level. The majority of the casework samples analyzed showed complete or nearly complete MH profiles even in degraded samples. To assess the informative power of MH profiles in forensic casework, probabilistic genotyping on partial MH profiles has been used. The resulting likelihood ratio values range from 7.84E+09 to 2.70E+34, thus defining an extremely strong support for the hypothesis that the genetic profile in a casework sample comes from the reference sample. Pairwise kinship simulations using allele frequencies from Italian population samples showed that full- and half-sibling relationships can be readily distinguished from unrelated individuals. For evaluation of the 76 MH panel’s utility for ancestry informativeness, PCA and STRUCTURE analyses are also presented comparing the newly collected sample from Ancona Italy with the 26 populations of the 1000 Genomes Project.</div><div>The results of the analysis confirmed the effectiveness of these short microhaplotypes in typing, with high sensitivity, samples with highly degraded DNA typically encountered in forensic cases.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103391"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145575146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The first population scale compendium of complete mitochondrial genomes from Flanders, Belgium","authors":"Noah Gaens ,&nbsp;Erika Souche ,&nbsp;Toomas Kivisild ,&nbsp;Jan Geypen ,&nbsp;Sara Seneca ,&nbsp;Francesc Calafell ,&nbsp;Maarten H.D. Larmuseau","doi":"10.1016/j.fsigen.2025.103382","DOIUrl":"10.1016/j.fsigen.2025.103382","url":null,"abstract":"<div><div>Mitochondrial DNA (mtDNA) plays a crucial role in forensics and population genetics, especially for analysing highly degraded samples. Yet, most mitogenomic reference datasets are still limited to control region (CR) sequences, that have limited discriminatory power. This is particularly true for Belgium, where despite the introduction of new legislation in 2024 to increase the use of mitochondrial DNA in familial searching, comprehensive mitogenomic reference data remain scarce. To fill this gap, we present a new forensic-grade dataset comprising complete mitochondrial genomes from 264 individuals with maternal ancestry from Flanders, the northern part of Belgium. Maternal lineages were verified through validated genealogical records extending prior to 1750. Our Flemish dataset reveals 252 unique haplotypes, a mean pairwise difference of 27.3, and a random match probability of 0.0042, demonstrating forensic resolution comparable to other high-quality European datasets. Haplogroups H, U, and T predominate in Flanders, aligning with broadly observed European patterns, while clustering analyses show close genetic affinities with neighbouring Western European populations. Furthermore, comparison with ancient genomes from the Middle Ages reveals notable continuity in maternal ancestry in Flanders throughout the historical era. Our data substantially expand the mitochondrial reference for Flanders, providing a high-quality resource based on autochthonous maternal lineages, suitable for forensic applications, genetic genealogical analyses, and microgeographic population studies.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103382"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145423655","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}