Forensic Science International-Genetics最新文献

{"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":"2025-09-26","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":"Salience, legitimacy, and credibility of two end-to-end forensic single cell probabilistic systems","authors":"Qhawe A. Bhembe ,&nbsp;Desmond S. Lun ,&nbsp;Ken R. Duffy ,&nbsp;Catherine M. Grgicak","doi":"10.1016/j.fsigen.2025.103369","DOIUrl":"10.1016/j.fsigen.2025.103369","url":null,"abstract":"<div><div>In cases for which there is no suspect, national forensic databases provide a mechanism by which to generate investigatory leads. National forensic DNA databases, however, have restrictions on what data to load. For example, uploading inferred alleles from DNA data that is a mixture of more than two contributors may be disallowed, leading to unresolved cases. A single-cell strategy has the potential to overcome the mixture gap by isolating each cell at the front-end of the pipeline. Once DNA signatures from each cell are obtained, they are clustered into groups. This is followed by asserting the probability we observe the data in the cluster had a person carrying genotype <em>g</em> donated. On applying Bayes’ Rule, we obtain the probability of a genotype given the data in a cluster and model. If this probability is near one, it means that only one genotype reasonably explains the data and this genotype can be used in a national database query. Good clustering, therefore, is an invaluable step in single-cell forensic interpretation and it is for this reason we examine the fortitude of two clustering approaches – i.e., model-based clustering (MBC) and forensic-aware clustering (FAC) – within an end-to-end single-cell predictor named EESCIt™. Using proper scoring rules, we report the performance of our probabilistic single-cell evaluator and structure the analytics into categories of <em>Salience</em>, <em>Legitimacy</em> and <em>Credibility</em> (SLC). With <em>Salience</em> referring to the applicability of a technology to meet an actor’s needs, we begin by discussing the relevance of single cell reports to forensic actors. Regarding <em>Legitimacy</em>, we determined the proportion of admixtures giving correct and incorrect cluster numbers and found that FAC returned correct cluster numbers for all admixtures tested. With improved clustering, 90 % of the loci returned only one credible genotype and it was the correct one, which improves on MBC’s 84 %. We then examined the Brier Score and decomposed it into calibration and refinement. We show that the FAC-centered system returned better calibration scores than the MBC one, which was driven by its improved clustering performance. Regarding <em>Credibility</em>, we found that the FAC-based system also returned better refinement scores. With FAC being more <em>Legitimate</em> and <em>Credible</em> than an MBC system for single-cell forensics, we adopt it into EESCIt™, therein creating the first end-to-end single-cell probabilistic system able to address single-cell queries about <em>how many</em> donors there were, and <em>who</em> they were.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103369"},"PeriodicalIF":3.1,"publicationDate":"2025-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145226832","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":"Nanopore sequencing and haplotyping of mitochondrial DNA hypervariable regions and its application on mixed stain","authors":"Yong Liao ,&nbsp;Junrui Shui ,&nbsp;Wenjie Chai ,&nbsp;Yaozhong Zou ,&nbsp;Chunying Jiang ,&nbsp;Huan Hu ,&nbsp;Yanan Liu","doi":"10.1016/j.fsigen.2025.103368","DOIUrl":"10.1016/j.fsigen.2025.103368","url":null,"abstract":"<div><h3>Objective</h3><div>To explore the feasibility of mitochondrial DNA (mtDNA) haplotype detection by nanopore sequencing and its potential value in the identification of mixed biological samples in forensic DNA analysis.</div></div><div><h3>Methods</h3><div>Firstly, validation of the method for long fragments detection of human mtDNA hypervariable regions (HVRs) was carried out on the G-seq500 nanopore sequencing platform. Then, the amplified products of two DNA standards were mixed at different ratios to be sequenced and analyzed. Finally, the method established in this study was used to separate haplotypes of the mixed biological samples from simulated and real crime scenes.</div></div><div><h3>Results</h3><div>The nanopore sequencing protocol of mtDNA HVRs established in this study demonstrated human-specificity, accuracy comparable to Sanger sequencing and Next Generation Sequencing (NGS), high sensitivity, repeatability and broad applicability. Using Sanger sequencing results as the standard, the accuracy of DNA standards sequencing was 100 %. The sequencing results for random individual blood cards were consistent with that of NGS. The detection limit of DNA template could be as low as 0.0625 ng. The method was also applicable to various sample types, including blood, oral swab, hair with follicles, different tissues and exfoliated cells, and can effectively interpret the Poly-C region. The mixtures of amplified products at ratios of 1:1, 1:9, 1:19, 1:49 and 1:99 from two DNA standards, as well as amplified products of mixed blood samples from two unrelated individuals at ratios of 3:2, 17:3, and 24:1, were accurately sequenced and haplotype-separated using the G-seq500 nanopore sequencing platform. Using this technical scheme to detect the mixture of exfoliated cells in a rape case can accurately separate the haplotypes to identify the suspect.</div></div><div><h3>Conclusion</h3><div>The long-read nanopore sequencing of human mtDNA HVRs based on the G-seq500 platform meets forensic DNA methodological requirements and is operationally feasible. It holds great potential and application value in the identification of mixed biological samples in forensic DNA analysis.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103368"},"PeriodicalIF":3.1,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145214774","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":"EFMex: Using EuroForMix to evaluate DNA mixtures with multiple persons of interest","authors":"Øyvind Bleka,&nbsp;Magnus Dehli Vigeland,&nbsp;Peter Gill","doi":"10.1016/j.fsigen.2025.103365","DOIUrl":"10.1016/j.fsigen.2025.103365","url":null,"abstract":"<div><div>We present EFMex (EuroForMix–Exhaustive), an R package for calculating likelihood ratios (LRs) in DNA mixture interpretation involving multiple persons of interest (POIs). The software implements an exhaustive method framework, evaluating all possible subsets of POIs as contributors to the mixture. We demonstrate the importance of this framework, particularly when contributors, such as close relatives, exhibit high allele sharing. A series of simulation experiments, designed to reflect complex casework scenarios, were conducted using three- and four-person mixtures in which at least two contributors were in the same family. The exhaustive method proved effective in distinguishing true contributors from closely related non-contributors, even under challenging conditions. A recalculation of exhaustive LR for candidates obtaining LR&gt;1 with the exhaustive method was used to increase discrimination further. An alternative to the exhaustive method, using a generalized likelihood ratio, returned values very close to those of the exhaustive method. A Shiny app was created as a graphical user interface to ease the analysis for practitioners.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103365"},"PeriodicalIF":3.1,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145118614","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":"Improving DNA detection sensitivity for the white-bellied pangolin (Phataginus tricuspis) via digital PCR","authors":"Georgia Kate Moloney ,&nbsp;Sean P. Heighton ,&nbsp;Nathalie Parthuisot ,&nbsp;Alain Didier Missoup ,&nbsp;Brice Roxan Momboua ,&nbsp;Chabi A.M.S. Djagoun ,&nbsp;Sery Gonedele-Bi ,&nbsp;Anne-Lise Chaber ,&nbsp;Philippe Gaubert","doi":"10.1016/j.fsigen.2025.103366","DOIUrl":"10.1016/j.fsigen.2025.103366","url":null,"abstract":"<div><div>Pangolins are widely regarded as the world’s most trafficked mammals, with their populations plummeting due to intense poaching to supplement the illegal trade in their scales, meat, and other derivatives. A persistent limitation in accurate trade tracing and successful criminal prosecution is the poor quality of seized specimens available for analysis, requiring the development of sensitive detection methodologies. Here, we developed a specific mtDNA-based digital PCR (dPCR) assay for the highly trafficked African white-bellied pangolin (<em>Phataginus tricuspis</em>) and applied it to a wide range of sample types. DNA extract concentration and quality, as assessed via fluorometry and 260/230–260/280 nm absorbance ratios, were highly heterogeneous among sample types due to differences in their inherent biological characteristics. To maximise the sensitivity of the dPCR assay, we used a ten-fold dilution series, resulting in an optimal dilution factor of 1/1000 applicable to any sample type. Due to cross-reactivity in the sister-species samples (<em>P. tetradactyla</em>), we developed a two-fold strategy to define assay detection limits based on the proportion of positive partitions (PPP). This resulted in a conservative threshold (PPP &gt; 0.00012, approximately 0.16 copies/µL) for <em>Phataginus</em> DNA detection, appropriate for general trade investigations, and a species-specific threshold (PPP &gt; 0.004, approximately 4.7 copies/µL) for the specific detection of <em>P. tricuspis</em>. Our success rate in identifying analysed <em>P. tricuspis</em> samples based on these thresholds was 80–70%, respectively. Negative dPCR outputs were significantly associated with lower DNA concentrations and poor-quality samples deviating from ideal absorbance ratio ranges, hence we recommend assessment of DNA quality indicators prior to dPCR testing to ensure optimal resource allocation. Levels of target DNA were variable among sample types. The positive correlation between DNA concentration and PPP indicated high levels of host-specific DNA in (i) rectal swabs, which were overall the best quality sample assessed, and (ii) bone and scale samples, which are the main materials available for trade scenarios. We present, to our knowledge, the first dPCR assay developed for pangolins, thus adding to the forensic toolkit available to curb their illegal trade. Despite our precautionary dPCR threshold estimation, we recommend that future studies reevaluate detection limits using available sample types and dilution series testing, while in the near future a multiplexing assay for all species seems reachable.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103366"},"PeriodicalIF":3.1,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145208641","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":"Characterisation of the UK domestic dog (Canis lupus familiaris) population for identification purposes using a forensically validated set of 13 autosomal STR markers","authors":"Federica Giangasparo ,&nbsp;David Ballard ,&nbsp;Ryan Colligan ,&nbsp;Florence Anderson ,&nbsp;Chiao-Yi Chung ,&nbsp;Brian Catchpole ,&nbsp;Denise Syndercombe Court","doi":"10.1016/j.fsigen.2025.103364","DOIUrl":"10.1016/j.fsigen.2025.103364","url":null,"abstract":"<div><div>Canine evidence in the UK is currently not being fully investigated, partly due to the lack of comprehensive genetic databases that are able to clarify the complexity of the highly inbred breeds that make up the canine population. The study here proposes the use of the CaDNAP markers, a forensically validated panel, to be applied to the wider UK canine population. For this purpose, over 1200 dogs have been genotyped for 13 STRs, belonging to 80 breeds. Mathematical corrections have been investigated to address the uneven breed distribution in the country in order to produce a more reliable allele frequency database for the whole UK canine population. In addition, a selection of the most common breeds (11) has been further investigated to produce allele frequency databases that are breed-specific with the aim of producing more accurate match probabilities should the breed of the animal of interest be known. The application of either set of frequencies has been scrutinized to evaluate their applicability in the field and are compared to current databases available in mainland Europe. A recommendation is made to use a minimum <em>F</em>_<em>ST</em> correction of 0.2 when estimating direct match likelihood ratios from whole country allele frequency databases.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103364"},"PeriodicalIF":3.1,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145109766","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":"Addendum to “Comparing six commercial autosomal STR kits in a large Dutch population sample”","authors":"Thirsa Kraaijenbrink","doi":"10.1016/j.fsigen.2025.103363","DOIUrl":"10.1016/j.fsigen.2025.103363","url":null,"abstract":"","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103363"},"PeriodicalIF":3.1,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145093284","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":"2025-09-09","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":"COMBO: An AmpliSeq-based trilogy for autosomal and uniparental (Y & mito) biogeographical ancestry and appearance testing","authors":"Christina Amory ,&nbsp;Robert Lagacé ,&nbsp;Walther Parson ,&nbsp;Catarina Xavier","doi":"10.1016/j.fsigen.2025.103349","DOIUrl":"10.1016/j.fsigen.2025.103349","url":null,"abstract":"<div><div>Forensic DNA phenotyping has become an increasingly important technology for narrowing down potential stain donors in forensic investigations. Over the past decade, the number of genetic markers required for reliable predictions has steadily increased, posing significant challenges for molecular genetic assay designs. These parallel designs often consume valuable DNA extracted from forensically relevant materials. To minimize DNA wastage, it is crucial to integrate as many markers as possible into a single analytical approach. Massively Parallel Sequencing (MPS)-based technologies have enabled the simultaneous analysis of up to several hundred genetic markers in one reaction. However, to date, marker panels from nuclear and mitochondrial DNA have not been integrated. This limitation has led to increased consumption of evidentiary DNA, as parallel analytical schemes are required to analyze both types of markers. This study presents the development, optimization and validation of the COMBO Panel, which combines autosomal and uniparental (Y-chromosomal and mitochondrial DNA) markers for ancestry and appearance testing. The COMBO panel contains 524 (VISAGE Enhanced Tool, ET) or 153 (VISAGE Basic Tool, BT) markers for the prediction of appearance and ancestry with 781 Y-chromosomal SNPs and 162 mtDNA amplicons covering the entire mitogenome. Library generation is based on AmpliSeq technology, which allows simultaneous analysis of all markers in a single sequencing run. The VISAGE ET panel was used in the optimization phase as well as in the initial trials of the COMBO assay and was later replaced with the VISAGE BT panel to streamline the test and adjust the primer concentrations accordingly. The COMBO panel was analyzed using Ion GeneStudio S5 systems for testing sensitivity and reproducibility, sex and species specificity, mock and real casework, degraded DNA and historical samples.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103349"},"PeriodicalIF":3.1,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144920335","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 coupling of methylation-sensitive restriction enzyme with recombinant polymerase amplification and lateral flow dipstick for the detection of (menstrual) blood stains","authors":"Xuan Tang ,&nbsp;Dan Wen ,&nbsp;Chudong Wang ,&nbsp;Hongtao Jia ,&nbsp;Yi Liu ,&nbsp;Xiaoyi Fu ,&nbsp;Bin Liang ,&nbsp;Jienan Li ,&nbsp;Hongzhe Wang ,&nbsp;Ying Liu ,&nbsp;Xingchun Zhao ,&nbsp;Lagabaiyila Zha","doi":"10.1016/j.fsigen.2025.103350","DOIUrl":"10.1016/j.fsigen.2025.103350","url":null,"abstract":"<div><div>Blood is a critical and frequently encountered type of evidence at forensic crime scenes. Its detection can provide valuable insights into the nature of a case and help refine investigative focus. The blood test strip is the most commonly used method for blood detection. However, it is prone to false-negative results, especially when detecting trace amounts of blood found at crime scenes, as well as challenging samples, such as those buried in soil, washed with detergents, and aged samples. In this study, we developed an alternative (menstrual) blood detection method based on the blood-specific methylation site (cg04011671). After treatment with a Methylation-Sensitive Restriction Enzyme (MSRE), the DNA is then amplified using Recombinant Polymerase Amplification (RPA), and the amplification is visualized through Lateral Flow Dipstick (LFD). Our results demonstrate that the MSRE-RPA-LFC method exhibits good specificity for body fluids and species. Additionally, this method shows a sensitivity of 125 pg DNA input, indicating its high detection capability for trace samples. Furthermore, this method is compatible with subsequent DNA analysis and demonstrates superior detection performance compared to traditional blood test strips (e.g., FOB test strips). It can effectively detect samples buried in soil for two weeks or longer, samples cleaned with reagents (e.g. hydrogen peroxide or oxygen bleach), and aged samples stored at room temperature for 10, 9, and 6 years. These findings indicate that this method serves as a confirmatory approach for (menstrual) blood identification and is a promising technique for detecting (menstrual) blood in trace and challenging forensic samples.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103350"},"PeriodicalIF":3.1,"publicationDate":"2025-08-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144908378","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}