Forensic Science International-Genetics最新文献

{"title":"Assessing the value of bacteria, plants, fungi and arthropods characterized via DNA metabarcoding for separation of forensic-like surface soils at varied spatial scales","authors":"Kelly A. Meiklejohn ,&nbsp;Melissa K.R. Scheible ,&nbsp;Tiffany Layne ,&nbsp;Teresa M. Tiedge ,&nbsp;Kim Love ,&nbsp;Jack Hietpas ,&nbsp;Jodi B. Webb ,&nbsp;Libby A. Stern","doi":"10.1016/j.fsigen.2025.103379","DOIUrl":"10.1016/j.fsigen.2025.103379","url":null,"abstract":"<div><div>Geological materials, including soil and dust, are ubiquitous and often inadvertently transferred during crime events. Forensic geologists use a range of particle-based analytical approaches to characterize the inorganic fraction for sample-to-sample comparisons. While analysis of the inorganic component often provides sufficient information to answer case related questions, there are inevitably cases where analysis of the organic fraction could be beneficial. In this study we assessed whether the biological communities associated with the organic fraction of North Carolina surface soils could allow for sample separation at four spatial scales: 1) sub-site samples – three samples collected 1 m apart, 2) small – A and B sites at a single location in close proximity (median distance of 66 m apart), 3) moderate – multiple locations within the same research station (median distance of 1.1 km apart), and 4) large – multiple locations within each physiographic region (Coastal Plain, Mountains, Piedmont) separated by up to 250 km. DNA metabarcoding coupled with Illumina® sequencing was used to amplify ∼200 bp regions of the following in each surface soil sample (n = 180): 1) 16S for bacteria, 2) ITS1 for fungi, 3) ITS2 and p6 loop of the <em>trnL-UAA</em> intron for plants, and 4) <em>COI</em> for arthropods. Generated sequences were taxonomically classified and subsequently organized into eight taxonomic combinations each analyzed at four data levels (unique sequence level and family rank, with and without removal of the 25 % most common taxa). Beta diversity was used as input for ANOSIM and PCA analyses to determine the optimal biological communities and data level to permit separation across the four spatial scales. The biological communities that permitted separation between samples varied based on the spatial scale being examined. At small spatial scales, datasets that included bacteria had the highest success of separating samples. At moderate spatial scales, the combination of plants + fungi or plants alone provided the highest success of separation. Finally, at the examined largest spatial scale, all combinations of biological communities tested performed comparably, likely due to the large diversity in habitat, land-use and geologic parent material between locations. Across all examined spatial scales, the optimal biological community combinations performed best when analyzed at the most granular unique sequence level. The results of this study demonstrate the value of biological communities for differentiating soils across varying spatial scales, providing a complementary analysis method for samples where the inorganic fraction lacked exclusionary differences.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103379"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145403244","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":"Bioinformatic processing of whole genome sequencing data with Tapir","authors":"August E. Woerner ,&nbsp;Briana N. Stowe ,&nbsp;Jonathan L. King ,&nbsp;Benjamin Crysup ,&nbsp;Meng Huang ,&nbsp;Michael D. Coble","doi":"10.1016/j.fsigen.2025.103387","DOIUrl":"10.1016/j.fsigen.2025.103387","url":null,"abstract":"<div><div>Whole genome sequencing (WGS) has powerful potential to aid forensic investigations. The scale and complexity of WGS, however, present sizable barriers to its widespread adoption. In particular, some form of standardized pipeline is needed to provide certainty in and an accounting of the final genotypes. To that end, we introduce Tapir (two-step analysis pipeline for investigative reporting). Tapir is a reproducible end-to-end scientific workflow that ingests raw WGS data from Illumina platforms (i.e., BCL format) and produces a GEDmatch-compatible genotyping result. Tapir combines many extant (and some custom) tools for forensic assessments of WGS data, including two modern and powerful probabilistic genotyping algorithms: one that relies on genotype refinement/imputation (GLIMPSE) and another that uses maximum likelihood (BCFtools). Additionally, Tapir provides summary and inferential statistics relevant to the forensic audience (e.g., breadth and depth of coverage, estimation of mixture status). Tapir comes with both mamba- and conda-compatible YAML environment files and has been packaged in a virtual machine image that allows it to run on commodity x86-64 computers, including Microsoft Windows.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103387"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145527923","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":"Multi-kingdom microbiota dynamics in exposed saliva: Integrating relative and absolute abundance across bacteria and fungi","authors":"Linying Ye ,&nbsp;Jiaqian Le ,&nbsp;Jieyu Du ,&nbsp;Litao Huang ,&nbsp;Weibin Wu ,&nbsp;Mingyue Zhao ,&nbsp;Quyi Xu ,&nbsp;Changhui Liu ,&nbsp;Chengliang Yang ,&nbsp;Chao Liu ,&nbsp;Ling Chen","doi":"10.1016/j.fsigen.2025.103390","DOIUrl":"10.1016/j.fsigen.2025.103390","url":null,"abstract":"<div><div>As an important body fluid of the human body, saliva harbors a diverse microbial community, yet the dynamic succession of its microbiota following environmental exposure remains incompletely understood. Most current studies focus solely on relative abundance and community structure derived from sequencing, which may lead to misinterpretation of microbial dynamics. Moreover, existing research has almost exclusively centered on bacterial communities, largely overlooking the role of fungi. In this study, we conducted a 90-day exposure experiment in two environments (an incubator and an indoor setting) and used 16S rRNA gene and ITS absolute quantification sequencing to characterize the structure and abundance of bacterial and fungal communities. Despite prolonged environmental exposure, core bacterial genera in saliva, such as <em>Streptococcus</em>, <em>Actinomyces</em>, <em>Rothia</em>, <em>Neisseria</em>, <em>Veillonella</em>, and <em>Prevotella</em>, persisted in the samples. An interaction between salivary and environmental bacterial communities was observed, which strengthened over time. Relative and absolute abundance analyses revealed different patterns: relative abundance suggested a moderately stable community structure with individual specificity, whereas absolute quantification demonstrated a significant decrease in total bacterial load over time. Fungal communities were nearly undetectable in fresh saliva but became detectable after 25 days of exposure in the incubator and subsequently increased in abundance, dominated by <em>Aspergillus</em>. In contrast to the significant decline in bacterial absolute abundance over time, fungi exhibited a continuous increase, with a negative correlation between the two. Relative abundance was more effective for individual identification, while absolute abundance performed better in time since deposition (TsD) estimation. Further incorporation of fungal data enhanced the accuracy of the TsD prediction model, as indicated by a mean absolute error (MAE) of 7.485 days and an R² of 0.905, highlighting the advantage of multi-kingdom microbial analysis.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103390"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145528322","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":"2026-02-01","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":"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":"2026-02-01","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":"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":"2026-02-01","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":"Genetic testing after sudden death with negative ancillary investigations: A French prospective study with multidisciplinary collaboration","authors":"Julien Osouf ,&nbsp;Audrey Schalk ,&nbsp;Elise Schaefer ,&nbsp;Nadège Calmels ,&nbsp;Angela Gonzalez ,&nbsp;Marie Claire Tortel ,&nbsp;Pascal Kintz ,&nbsp;Jean-Sébastien Raul ,&nbsp;Caroline Schulth-Bolard ,&nbsp;Audrey Farrugia","doi":"10.1016/j.fsigen.2025.103375","DOIUrl":"10.1016/j.fsigen.2025.103375","url":null,"abstract":"<div><h3>Background</h3><div>In case of unexplained sudden death, learned societies recommend performing genetic testing on post-mortem biological specimens to search for a potential hereditary genetic cause and implement, with a multidisciplinary team, screening and prevention measures for the deceased's relatives. The aim of this study was to assess the diagnostic contribution of a rigorous multidisciplinary approach (combining autopsy, toxicology, anatomopathology and genetics) in case of unexplained sudden death.</div></div><div><h3>Methods</h3><div>This was a French, prospective (2008–2020), monocentric cohort study on unexplained sudden death in young subjects (aged 2–50 years). For each case, a rigorous autopsy, and the analysis of anatomopathological, toxicological, and genetic data were performed. Fresh cardiac and brain tissue samples were systematically collected and stored at −80°C. Genetic testing was performed using a targeted panel of 181 genes associated with channelopathies, cardiomyopathies and epilepsy. In negative cases, whole exome sequencing was performed. Exomes analyzed more than two years earlier were also reanalyzed.</div></div><div><h3>Results</h3><div>One or two pathogenic or likely pathogenic (P/LP) variants were identified in 23.5 % (8/34) of patients, and positive toxicology results as well as notable microscopic cardiac anomalies were identified in 25 % (2/8) of them. One or two variants of uncertain significance (VUS) were identified in 35 % (12/34) of patients, of whom 50 % (6/12) had positive toxicology results and 58.3 % (7/12) showed microscopic cardiac anomalies. In addition, 2.9 % (1/34) of patients carried both a pathogenic variant and a VUS in another gene. The identified variants were associated with cardiac, neurological or metabolic disorders, or found in a gene of uncertain significance. Two patients carried pathogenic variants in genes with an uncertain association with sudden death. A total of 20 different genes carried variants of interest (VUS, P/LP variant): <em>KCNH2, SCN5A, RYR2, SNTA1, PRKAG2, AKAP9, GDP1L, TECRL, TRDN, HCN4, ZFHX3, DEPDC5, TANC2, CHRNA2, DCHS1, DMD, EYA4, ACADM, KLF10, SCN7A.</em></div></div><div><h3>Conclusion</h3><div>Our study highlighted the value of extensive genetic testing in case of unexplained sudden death, reinforcing the importance of using a comprehensive, multidisciplinary approach. The presence of multiple genetic variants, minor anatomopathological changes and toxicological factors suggests the complex interplay of contributory mechanisms leading to death. Regular reanalysis of genetic data, combined with close collaboration between forensic pathologists, geneticists and clinicians, is essential to improve diagnostic yield and guide family management.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103375"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145350610","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":"2026-02-01","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":"Development and validation of an age prediction model using mRNA markers","authors":"Mengxiao Liao ,&nbsp;Anqi Chen ,&nbsp;Yujia Xuan ,&nbsp;Jinyuan Zhao ,&nbsp;Sitong Liu ,&nbsp;Yu Xing ,&nbsp;Suhua Zhang ,&nbsp;Chengtao Li","doi":"10.1016/j.fsigen.2025.103373","DOIUrl":"10.1016/j.fsigen.2025.103373","url":null,"abstract":"<div><div>As a downstream product of transcriptional regulation, mRNA offers valuable insights into age-associated molecular changes and shows considerable potential for applications in age estimation. In this study, RNA sequencing was conducted on peripheral blood samples from 127 healthy Chinese individuals spanning a broad age range (18–80 years). Differential expression analysis were performed on expression profiles among the young (＜30 years), middle-aged (30–60 years) and elderly (＞60 years) groups. A total of 79 differentially expressed genes (DEGs) were identified. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis revealed that the DEGs were mainly involved in response to type I interferon and cell-cell adhesion, which may reflect the activation of pro-inflammatory responses and neurodegenerative changes associated with aging. Through Spearman correlation analysis and the Lasso regression method, 34 candidate age-related genes (ARGs) were selected as predictive features. Notably, seven of these mRNAs (ARHGEF4, ARF6, AMIGO1, FITM2, PLEKHG4, SLC5A10, and HKR1) were identified for the first time as ARGs. Age prediction models were then constructed using five machine learning algorithms, with a 7:3 split for the train set and test set. Internal validation was conducted using 20 samples from the same cohort, while external validation was performed using two independent datasets: GSE262619 and GSE124326. Among them, the elastic net model showed the best performance, yielding a mean absolute error (MAE) of 6.72 years on the test set, 7.56 years on the internal validation set and 11.74 years on the external validation set, demonstrating strong robustness in age prediction. In general, this study delineated age-associated mRNA expression patterns and established a robust and accurate age prediction model based on a minimal set of 34 mRNA markers.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"81 ","pages":"Article 103373"},"PeriodicalIF":3.1,"publicationDate":"2026-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145331494","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":"2026-02-01","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}