This study explores the development and evaluation of miRNA and mRNA panels for body fluid identification.
This project explores the use of microRNA (miRNA) and messenger RNA (mRNA) for body fluid identification (BFID). The results provide the forensic community with information on the performance of miRNA and mRNA for BFID using capillary electrophoresis (CE), quantitative reverse transcription PCR (RT-qPCR), and massively parallel sequencing (MPS) technologies, particularly for challenging samples. In the first study, an eight-marker miRNA multiplex was developed for capillary electrophoresis using a linear primer system. In the second study, the stability and persistence of miRNA and mRNA was evaluated. Custom reverse transcription quantitative PCR (RT-qPCR) assays were performed. A persistence study was also performed by laundering blood and semen-stained swatches and either machine drying or allowing the samples to air dry. In the final study, an early access mRNA panel for BFID was evaluated. Reverse transcription negatives, total reads, and composition of reads attributed to body fluid specific markers were used to evaluate the performance of the panel.