{"title":"A CE-based mRNA profiling method including six targets to estimate the time since deposition of blood stains","authors":"A.E. Fonneløp , N.V. Hänggi , C.C. Derevlean , Ø. Bleka , C. Haas","doi":"10.1016/j.fsigen.2025.103240","DOIUrl":null,"url":null,"abstract":"<div><div>An association between RNA degradation and the time since deposition (TsD) of a biological stain has previously been demonstrated. Despite the encouraging results obtained with several RNA markers, the variability in results between individuals and analytical approaches limits the method's application in casework. The incorporation of multiple markers into a single prediction model could enhance estimation accuracy. Typically, real-time qPCR has been the primary analytical platform for these studies. However, qPCR requires high sample volumes and involves numerous pipetting steps when analysing multiple markers, increasing the risk of errors. In this study, we aim to optimize the TsD analysis by combining six targets in three mRNA markers (S100A12, LGALS2 and CLC) in a PCR multiplex and transitioning the analysis platform from qPCR to capillary electrophoresis (CE). This collaborative effort between the Department of Forensic Research at Oslo University Hospital (Laboratory 1) and the Zurich Institute of Forensic Medicine (ZIFM, Laboratory 2) analysed a total of six sample sets, spanning a period of 0 days up to 1.5 years (551 days), along with a broad set of test samples including different carrier materials. Furthermore, a machine learning model was employed to predict the age of bloodstains, aiming to enhance the precision and reliability of TsD estimations.</div></div>","PeriodicalId":50435,"journal":{"name":"Forensic Science International-Genetics","volume":"77 ","pages":"Article 103240"},"PeriodicalIF":3.2000,"publicationDate":"2025-02-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Forensic Science International-Genetics","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1872497325000201","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
引用次数: 0
Abstract
An association between RNA degradation and the time since deposition (TsD) of a biological stain has previously been demonstrated. Despite the encouraging results obtained with several RNA markers, the variability in results between individuals and analytical approaches limits the method's application in casework. The incorporation of multiple markers into a single prediction model could enhance estimation accuracy. Typically, real-time qPCR has been the primary analytical platform for these studies. However, qPCR requires high sample volumes and involves numerous pipetting steps when analysing multiple markers, increasing the risk of errors. In this study, we aim to optimize the TsD analysis by combining six targets in three mRNA markers (S100A12, LGALS2 and CLC) in a PCR multiplex and transitioning the analysis platform from qPCR to capillary electrophoresis (CE). This collaborative effort between the Department of Forensic Research at Oslo University Hospital (Laboratory 1) and the Zurich Institute of Forensic Medicine (ZIFM, Laboratory 2) analysed a total of six sample sets, spanning a period of 0 days up to 1.5 years (551 days), along with a broad set of test samples including different carrier materials. Furthermore, a machine learning model was employed to predict the age of bloodstains, aiming to enhance the precision and reliability of TsD estimations.
期刊介绍:
Forensic Science International: Genetics is the premier journal in the field of Forensic Genetics. This branch of Forensic Science can be defined as the application of genetics to human and non-human material (in the sense of a science with the purpose of studying inherited characteristics for the analysis of inter- and intra-specific variations in populations) for the resolution of legal conflicts.
The scope of the journal includes:
Forensic applications of human polymorphism.
Testing of paternity and other family relationships, immigration cases, typing of biological stains and tissues from criminal casework, identification of human remains by DNA testing methodologies.
Description of human polymorphisms of forensic interest, with special interest in DNA polymorphisms.
Autosomal DNA polymorphisms, mini- and microsatellites (or short tandem repeats, STRs), single nucleotide polymorphisms (SNPs), X and Y chromosome polymorphisms, mtDNA polymorphisms, and any other type of DNA variation with potential forensic applications.
Non-human DNA polymorphisms for crime scene investigation.
Population genetics of human polymorphisms of forensic interest.
Population data, especially from DNA polymorphisms of interest for the solution of forensic problems.
DNA typing methodologies and strategies.
Biostatistical methods in forensic genetics.
Evaluation of DNA evidence in forensic problems (such as paternity or immigration cases, criminal casework, identification), classical and new statistical approaches.
Standards in forensic genetics.
Recommendations of regulatory bodies concerning methods, markers, interpretation or strategies or proposals for procedural or technical standards.
Quality control.
Quality control and quality assurance strategies, proficiency testing for DNA typing methodologies.
Criminal DNA databases.
Technical, legal and statistical issues.
General ethical and legal issues related to forensic genetics.