COMPARATIVE EVALUATION OF METHODS FOR DIFFERENTIATING BIOLOGICAL FLUIDS BASED ON MRNA PROFILES
Abstract
(1) Background: Accurate identification of biological fluids at crime scenes is a cornerstone of forensic molecular biology. Traditional presumptive and confirmatory tests are often limited in specificity and sensitivity, and mRNA-based profiling has emerged as a powerful tissue-specific identification approach. (2) Methods: This review systematically compares four primary mRNA detection platforms — reverse transcription quantitative PCR (RT-qPCR), DNA microarray analysis, NanoString nCounter technology, and RNA sequencing (RNA-Seq) — applied to the differentiation of blood, saliva, semen, vaginal secretions, menstrual blood, and skin cells. Published validation studies and forensic case reports were analyzed. (3) Results: RT-qPCR demonstrated the best balance of sensitivity, specificity, and cost-effectiveness for routine forensic application. NanoString nCounter showed superior multiplexing capacity and reproducibility. RNA-Seq offered the highest resolution but remains impractical for most forensic laboratories due to cost. LAMP-based methods represent a promising rapid alternative. (4) Conclusions: mRNA profiling provides reliable, tissue-specific identification of biological fluids superior to conventional methods. RT-qPCR and NanoString platforms are currently best suited for forensic implementation, while RNA-Seq may become standard practice as costs decrease.
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