Forensic Science International-Genetics最新文献

{"title":"Long non-coding RNAs in human peripheral blood as biomarkers for age prediction","authors":"Anqi Chen ,&nbsp;Yujia Xuan ,&nbsp;Fan Yang ,&nbsp;Yanan Liu ,&nbsp;Jinyuan Zhao ,&nbsp;Mengxiao Liao ,&nbsp;Yanfang Lu ,&nbsp;Yu Xing ,&nbsp;Suhua Zhang ,&nbsp;Chengtao Li","doi":"10.1016/j.fsigen.2025.103419","DOIUrl":"10.1016/j.fsigen.2025.103419","url":null,"abstract":"<div><div>Chronological age is a critical biological trait with substantial relevance in forensic investigations. Growing evidence highlights the role of long non-coding RNAs (lncRNAs) in aging, but the application in age prediction remains largely unexplored. To evaluate the potential of lncRNAs for age prediction, we analyzed RNA-seq data from peripheral blood samples of Han Chinese individuals. Differential expression analysis identified a series of differentially expressed lncRNAs (DElncRNAs) among multiple age group comparisons, with the most significant expression changes observed between the youngest (18–27 years) and older (68–77 years) age groups. GO and KEGG enrichment analysis of target genes indicated that these lncRNAs were involved in mitochondrial function, telomere maintenance, and pathways related to neurodegenerative disease. Additionally, Spearman’s correlation analysis further identified 20 lncRNAs significantly associated with age as candidate markers, which were subsequently used to construct age prediction models using six machine learning algorithms. The XGBoost model achieved the best performance, with a mean absolute error (MAE) of 8.04 years (R² = 0.66) in the test cohort. External validation was conducted using two independent datasets (GSE124326 and GSE177044). The XGBoost and LightGBM models consistently outperformed the other models, with MAEs ranging from 7.99 to 8.21 years. These findings demonstrated the feasibility of lncRNA-based age prediction, providing novel biomarkers and methodological insights for forensic age estimation.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103419"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145938492","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":"Challenges in mRNA-based time since deposition estimation: A multiplex MPS primer panel investigation","authors":"Nadescha Viviane Hänggi ,&nbsp;Melisa Walliser ,&nbsp;Ane Elida Fonneløp ,&nbsp;Robert Lagacé ,&nbsp;Erin Hanson ,&nbsp;Jack Ballantyne ,&nbsp;Cordula Haas","doi":"10.1016/j.fsigen.2025.103415","DOIUrl":"10.1016/j.fsigen.2025.103415","url":null,"abstract":"<div><div>Estimating the time since deposition (TsD) of biological stains can provide crucial information for crime scene investigations, but remains a challenging task. In this study, we investigated mRNA degradation patterns across four forensically relevant body fluids - blood, semen, menstrual blood, and vaginal secretion - using a custom-designed mRNA MPS primer panel. The primer panel included candidate mRNA markers identified by RNA-Seq for TsD estimation in multiple body fluids. It was designed to assess mRNA degradation patterns based on three aspects: (1) individual mRNA degradation trends, (2) intra-transcript stability, examining 5′- versus 3’-end degradation, and (3) inter-transcript stability, based on relative degradation between two targets. The results revealed high inter-individual variability, especially for low-abundance transcripts. Some mRNA markers showed degradation patterns that correlated with TsD. However, inconsistencies in the degradation patterns observed across the analyzed time series highlight the challenges associated with primer panel optimization, RNA extraction methods, and reproducibility across samples. Our results suggest that, while mRNA degradation remains a promising strategy for TsD estimation, its forensic application requires careful marker selection and validation across independent data sets, to account for methodological and biological variability.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103415"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145919497","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":"LAMPSEQ: Colorimetric LAMP and nanopore sequencing for rapid species identification in the illegal wildlife trade","authors":"O. Yugovich ,&nbsp;S. Sturrock ,&nbsp;V. Cave ,&nbsp;S.A. Harbison","doi":"10.1016/j.fsigen.2025.103412","DOIUrl":"10.1016/j.fsigen.2025.103412","url":null,"abstract":"<div><div>The illegal ivory trade in animal parts such as ivory, horn, and bone, threatens the survival of many protected species including elephants, rhinoceros, hippopotamus, and walrus. Accurate and reliable species identification is critical for enforcement under CITES, yet traditional DNA-based methods are time-consuming, resource-intensive, and are often inaccessible in regions most affected by poaching. This proof-of-concept study presents a rapid, portable, two-step workflow combining colorimetric loop-mediated isothermal amplification (LAMP) with nanopore sequencing for the authentication of elephant ivory and its legal and illegal substitutes. Novel LAMP assays were developed for elephants, mammoths, rhinoceros, hippopotamus, walrus, water buffalo, and common domestic species, targeting conserved mitochondrial DNA regions to enable both presumptive detection and sequencing-based confirmation. Using synthetic DNA, all assays demonstrated high sensitivity, with limits of detection down to 1–100 fg, with good consistency between replicates. A multiplexing strategy was designed to group closely related species into a single assay. Separate assays were designed for all elephant species, all mammoths, and the two closely related rhinoceros species (<em>R. sondaicus</em> and <em>R. unicornis</em>), enabling rapid presumptive screening followed by species-level confirmation via sequencing. To address challenges associated with the tandem repeats present in LAMP products, LAMPSEQ, a custom bioinformatics pipeline, was developed to extract, align, and accurately identify species from LAMP sequencing data. This sensitive, scalable, and field-deployable workflow expands the molecular toolkit, delivering innovative solutions to wildlife crime investigations and supports the protection of the world’s most vulnerable and treasured species from extinction.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103412"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145797409","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":"ForSpEC: A compact forensic epigenetic age clock for sperm cells with cross‑platform validation","authors":"Aleksandra Pisarek-Pacek ,&nbsp;Joanna Rudnicka ,&nbsp;Rezvan Noroozi ,&nbsp;Bożena Wysocka ,&nbsp;Aleksander Masny ,&nbsp;Magdalena Spólnicka ,&nbsp;Andrzej Ossowski ,&nbsp;Aneta Sitek ,&nbsp;Aneta Macur ,&nbsp;Jarosław Janeczko ,&nbsp;Gabriela Kania ,&nbsp;Marta Sikora-Polaczek ,&nbsp;Wojciech Branicki ,&nbsp;Ewelina Pośpiech","doi":"10.1016/j.fsigen.2026.103426","DOIUrl":"10.1016/j.fsigen.2026.103426","url":null,"abstract":"<div><div>Expanding the use of forensic DNA intelligence requires the development of accurate DNA-based predictive models. The primary challenge in achieving a highly accurate epigenetic age prediction method in sperm lies in the unique biological function of male reproductive cells. In this study, we independently evaluated the Germ Line Age (GLA) calculator, one of the most comprehensive tools for predicting sperm age to date. We then set out to define a streamlined, well-curated set of sperm cell-specific markers suitable for forensic applications, while providing an integrated laboratory tool for targeted methylation profiling using bisulfite amplicon sequencing with Ion AmpliSeq™ technology. DNA was extracted from 212 sperm samples, genome-wide DNA methylation was measured using Infinium MethylationEPIC (EPIC) arrays and methylation beta values were extracted for 260 literature-reported CpG markers. Application of the GLA model to this sperm cell dataset resulted in a mean absolute error (MAE) of 2.82 years. As a next step, predictive analysis based on 147 training set samples identified a subset of eight optimal CpG markers for model building. Due to technical constraints encountered during Ion AmpliSeq™ assay optimization, the final model was refined to include seven CpGs, yielding the ForSpEC (<u>For</u>ensic <u>Sp</u>erm <u>E</u>pigenetic <u>C</u>lock) model. ForSpEC predicted age with an MAE of 3.13 years in the 65 test samples. When the EPIC-trained ForSpEC model was applied to Ion AmpliSeq™ data (N = 20), age prediction accuracy remained high, with an MAE of 3.76 years. For comparison, existing predictors were retrained using EPIC data, demonstrating that the seven CpGs comprising ForSpEC rank among the most accurate predictors of sperm epigenetic age. The model was further validated using data from publicly available repositories and the impact of cellular composition on prediction accuracy was additionally addressed by comparing DNA samples isolated from sperm cells with whole semen samples. Overall, this study advances sperm-based epigenetic age estimation by introducing ForSpEC, a highly accurate forensic epigenetic clock that relies on just seven CpG markers. This new method exhibits only a modest reduction in accuracy compared to the GLA model, which relies on 261 CpGs. Moreover, the analyses underscore the high utility of Ion AmpliSeq™ technology for targeted DNA methylation analysis in forensic applications.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103426"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146021030","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":"Construction and validation of a rapid semen identification system based on SHERLOCK technology","authors":"Yu Luo ,&nbsp;Xianmiao Wang ,&nbsp;Fan Yang ,&nbsp;Yixia Zhao ,&nbsp;Sheng Hu ,&nbsp;Shuyu Liu ,&nbsp;Shuang Li ,&nbsp;Guochang Luo ,&nbsp;Qifan Sun","doi":"10.1016/j.fsigen.2025.103410","DOIUrl":"10.1016/j.fsigen.2025.103410","url":null,"abstract":"<div><div>This study developed a rapid detection system for semen-specific mRNA based on CRISPR/Cas13a system to meet the timeliness requirements of forensic on-site body fluid identification. Specific primers and CRISPR RNA (crRNA) short fragments on semen specific mRNA genes were designed and screened, to establish a SHERLOCK detection method based on technology principles of CRISPR/Cas. Furthermore, nucleic acid rapid release agents for treating samples were screend to construct a new detection method in combination with SHERLOCK, and the specificity and sensitivity of the method were tested. The method can rapidly detect the presence of semen from unknown body fluid samples, and the relative fluorescence unit (RFU) value of the semen sample is significantly higher than those of non-semen samples (P &lt; 0.0001), with a sample detection sensitivity of down to 0.25 μL. The construction of the rapid semen detection method using rapid extraction and SHERLOCK reduces operation time, significantly reduces instrument dependence, and provides an innovative solution for forensic on-site rapid body fluid identification.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103410"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145746215","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":"Enhancing forensic casework interpretation through DNA methylation-based analysis: A case study of pooled blood samples","authors":"Jeong Min Lee ,&nbsp;Sang Un Park ,&nbsp;Bo Min Kim ,&nbsp;Hanbyeol Kim ,&nbsp;Hwan Young Lee","doi":"10.1016/j.fsigen.2026.103424","DOIUrl":"10.1016/j.fsigen.2026.103424","url":null,"abstract":"<div><div>In forensic casework, the meticulous investigation of unidentified biological material is critical for developing decisive investigative leads. Integrated genetic and epigenetic profiling allows for the reconstruction of incident scenarios and the assessment of criminal involvement probability. Specifically, DNA methylation-based analyses facilitate the determination of cellular composition, donor age, and smoking status, progressively delineating probable incident circumstances and converging on donor identification. We report a complex case involving a large volume of pooled blood discovered on an apartment staircase half-landing with an unknown donor identity. The investigation was refined using multiple genetic and epigenetic assays, including STR typing, Y-haplotyping, SNaPshot-based body fluid identification, and array-based DNA methylation profiling. Two samples collected at the scene—one presumed blood and one unknown—were confirmed to originate from the same donor: a Korean male belonging to Y-haplogroup O3a2. SNaPshot analysis indicated the presumed blood sample was a blood–saliva mixture, while the unknown sample remained unassigned to any specific fluid type. Array-based cell/tissue deconvolution and unsupervised clustering analysis further revealed that the presumed blood sample was predominantly composed of whole blood with a minor contribution from upper gastrointestinal tissue. Integrating the results of body fluid identification with cell-proportion deconvolution supported the interpretation of hematemesis (vomiting blood), thereby favoring a medical scenario over criminal involvement. The unknown sample exhibited methylation patterns distinct from all candidate tissue groups, possibly reflecting sample quality limitations or tissue mixture. Chronological epigenetic clock analyses estimated the donor’s age to be middle-aged (approximately 50–60 years), while smoking status prediction classified the donor as a current smoker. Despite suboptimal sample quality and the complete absence of contextual information, this comprehensive analysis provided detailed donor profiling and incident reconstruction. This case exemplifies the power of integrated genetic and epigenetic methodologies—particularly tissue-of-origin profiling—to generate investigative leads, facilitate the inference of incident circumstances, and provide substantive contributions to forensic investigations.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103424"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145977150","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":"Target RNA sequencing system for identification of body fluid contributors in mixtures","authors":"Huan Yu,&nbsp;Jiaxin Ji,&nbsp;Jiayan Li,&nbsp;Jiaqi Wang,&nbsp;Zidong Liu,&nbsp;Xiaoxiao Wang,&nbsp;Yiwei Guo,&nbsp;Liangze Li,&nbsp;Jinding Liu,&nbsp;Zeqin Li,&nbsp;Gengqian Zhang","doi":"10.1016/j.fsigen.2025.103416","DOIUrl":"10.1016/j.fsigen.2025.103416","url":null,"abstract":"<div><div>The cSNPs within body fluid-specific RNA molecules have emerged as a promising biomarker for body fluid identification and identifying body fluid donors in mixture stains. Unlike traditional DNA analysis, which is restricted to individualizing a forensic stain at a sub-source level, RNA-SNP profiling enables direct association of the body fluid with the suspect. However, the number of available RNA-SNPs is still not enough to achieve a satisfactory discriminatory power. To explore novel SNPs for RNA-based genotyping, we screened out a series of potential RNA markers and developed a target RNA sequencing assay comprising 135 RNA-SNPs across 42 body fluid genes to identify body fluid contributors in the mixture. The assessment of specificity was performed on single-source samples, and the results exhibited a clear separation between five body fluid types. The assay also showed complete genotype concordance between gDNA and cDNA, and had a capacity to analyze aged stains. In the artificial RNA mixtures consisting of 2–5 body fluid components and 2-component mock case mixtures, all body fluid components could be successfully detected, and most of the genotypes could be correctly associated with their body fluid contributors, indicating excellent discriminative capacity of our panel for mixture deconvolution. The panel presented a cumulative discriminatory power ranging from 0.987325116 (saliva) to 0.999993412 (venous blood), and could serve as an effective complementary tool to improve discriminatory power when combined with other currently available RNA-SNP panels.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103416"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145866411","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":"CRISPR/Cas12a coupled with MIRA: A specific and rapid assay for human DNA in challenging forensic matrices","authors":"Si-rui Li ,&nbsp;Yang Li ,&nbsp;Kai-bo Yang ,&nbsp;Si-wen Wang ,&nbsp;Mao-ling Sun ,&nbsp;Zhenze Liu ,&nbsp;Xiu-peng Zhang ,&nbsp;Yang Zhong ,&nbsp;Jun Yao","doi":"10.1016/j.fsigen.2025.103393","DOIUrl":"10.1016/j.fsigen.2025.103393","url":null,"abstract":"<div><div>Human DNA detection is crucial in forensic medicine, particularly for trace, degraded, or mixed samples, which demand high sensitivity, specificity, and rapid processing. Traditional methods, such as immunological assays and PCR-based techniques, often suffer from operational complexity, limited sensitivity, or high equipment dependency. To address these challenges, we developed a novel detection system combining multienzyme isothermal rapid amplification (MIRA) with CRISPR-Cas12a for the rapid, specific, and portable human DNA identification. By targeting the human mitochondrial cytochrome b (CYTB) gene and sex-determining Region Y(SRY) gene, we designed MIRA primers and CRISPR-Cas12a crRNA to enable dual recognition and signal amplification. The method involves isothermal amplification at 37°C followed by CRISPR-Cas12a-mediated cleavage, producing detectable fluorescence or lateral flow chromatographic signals. Our system achieves ultra-sensitive detection and high specificity, distinguishing human DNA from non-human sources (e.g., pig, chicken, mouse), and also enables accurate gender identification, further enhancing its utility in forensic and genetic studies. Compared to traditional qPCR, this approach demonstrates superior sensitivity, faster turnaround (≤ 45 min), and minimal equipment requirements, making it ideal for forensic applications. Moreover, the blood, mixed, and degraded samples were used to confirm its robustness, with results interpretable via blue-light fluorescence or colloidal gold test strips. In summary, the MIRA-CRISPR/Cas12a system overcomes the limitations of conventional techniques, offering a rapid, cost-effective, and reliable solution for forensic human DNA identification, with potential extensions to wildlife conservation and food safety testing.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103393"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145727721","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":"Forensic Lab-on-a-Disc for enhanced sperm cell purification in differential extraction workflows","authors":"Un Na Koh ,&nbsp;Vijaya Sunkara ,&nbsp;Beomhee Ahn ,&nbsp;Mi Kang Jung ,&nbsp;Myunghee Jang ,&nbsp;Hwanghee Ryu ,&nbsp;Hanna Kim ,&nbsp;Jihye Ahn ,&nbsp;Kyusang Lee ,&nbsp;Si-Keun Lim ,&nbsp;Yoon-Kyoung Cho ,&nbsp;Beomseok Lee","doi":"10.1016/j.fsigen.2026.103430","DOIUrl":"10.1016/j.fsigen.2026.103430","url":null,"abstract":"<div><div>Efficient separation of sperm cells from mixed biological samples is critical in forensic DNA analysis, particularly in sexual assault investigations. Sperm separation in conventional differential extraction (cDE) methods is labor-intensive, time-consuming, and prone to sample loss, often compromising DNA recovery and quality. Here, we present the Forensic Lab-on-a-Disc, a centrifugal microfluidic device that enables rapid, semi-automated sperm cell and non-sperm DNA separation. Leveraging centrifugal force and tangential flow filtration with dual membranes (10 µm and 1 µm pore sizes), the device effectively separates sperm cells from epithelial cell lysates and debris. Optimization experiments identified a 10 µm pore size for the first filter as optimal for maximizing sperm cell recovery while depleting non-sperm DNA. A single washing cycle was sufficient to preserve male DNA ratio (defined as Y-DNA divided by total gDNA) while minimizing sperm loss. Comparative analyses showed that the Forensic Lab-on-a-Disc, which specifically replaces the sperm-cell separation step within the cDE workflow, outperforms the conventional method by delivering significantly higher male DNA ratio, faster processing times (∼6 min), and improved reproducibility. STR profiling confirmed male DNA recovery with minimal non-sperm DNA carryover, even in samples with epithelial-to-sperm DNA equivalent ratios as high as 1000:1. Featuring a compact, semi-automated, and multi-sample processing capability, the Forensic Lab-on-a-Disc reduces operator variability and minimizes non-sperm–derived lysate carryover risks that compromise mixture interpretation, offering a transformative solution to accelerate forensic DNA analysis in sexual assault cases.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103430"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146069422","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 impact of illicit drugs on DNA extraction efficiency","authors":"Madison Nolan ,&nbsp;K. Paul Kirkbride ,&nbsp;Adrian Linacre","doi":"10.1016/j.fsigen.2025.103413","DOIUrl":"10.1016/j.fsigen.2025.103413","url":null,"abstract":"<div><div>The analysis of illicit drugs for the presence of human DNA provides an opportunity to gain greater information on ‘who’ is involved in the illicit drug supply chain, complementing the existing chemical profiling techniques to identify ‘what’ and ‘where’. Given the known inhibitory effect of numerous illicit drugs and related compounds on PCR, it is crucial to understand the effectiveness of currently used DNA extraction kits to successfully remove these compounds while extracting sufficient DNA for downstream DNA profiling. To do this, four common illicit drugs (methamphetamine HCl, heroin HCl, MDMA HCl and GHB Na) in addition to two precursors of methamphetamine (pseudoephedrine HCl and phenyl-2-propanone) were spiked with saliva and DNA extracted. Two DNA extraction kits were selected to evaluate columns compared to beads, the QIAGEN QIAamp® DNA Investigator kit and the Promega DNA IQ™ system. DNA quantification of these samples yielded significantly more DNA recovery for each drug, except P2P, when extracted using DNA Investigator. Similarly, peak heights and heterozygous peak height balance were, on average, improved when samples were extracted with DNA Investigator compared to DNA IQ. Regardless of drug or extraction kit, no inhibition was detected during DNA quantification or profiling, indicating reduced DNA yields were caused by inefficient DNA extraction rather than inhibition of the PCR processes. All samples extracted using DNA Investigator yielded complete DNA profiles while DNA IQ yielded near-complete profiles, ultimately resulting in extremely strong likelihood ratio values from all samples. The generation of informative DNA profiles, despite the presence of illicit drugs, using two commercially available DNA extraction kits highlights the potential to implement these sample types into operational laboratories.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"82 ","pages":"Article 103413"},"PeriodicalIF":3.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145829442","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}