P245 – Cross‑platform Validation of a mRNA Assay for Differentiating Menstrual Blood from Peripheral Blood

11:13

Thursday August 20^th

Station 17

Duration: 12 minutes

04. Forensic biology

Niu Gao

The accurate differentiation of menstrual blood from peripheral blood is of significant forensic importance, particularly in sexual assault and violent crime investigations involving female victims. Although mRNA profiling provides high tissue specificity, most existing assays have been validated on a single analytical platform and lack universally applicable data interpretation criteria across different platforms. In this study, we selected ten mRNA markers, including four menstrual blood-specific markers (MMP7, MMP10, SFRP4, STC1), four peripheral blood-specific markers (HBA, HBB, ALAS2, GYPA), and two housekeeping genes (ACTB, 18S rRNA), and simultaneously used three analytical platforms to evaluate the performance of these markers in distinguishing menstrual blood from peripheral blood. Identical primer sets were applied across quantitative real-time PCR (qPCR), capillary electrophoresis (CE), and massively parallel sequencing (MPS), enabling direct cross-platform comparison of specificity, sensitivity, and mixture performance. We further established interpretation criteria of test results to indicate the presence or absence of menstrual blood. Interestingly, results were largely consistent across qPCR, CE, and MPS: Peripheral blood-specific markers were detected in both menstrual and peripheral blood samples, whereas menstrual blood-specific markers were detected exclusively in menstrual blood; GYPA and SFRP4 exhibited occasional dropout in menstrual blood samples when the mRNA input was below 1 ng; Three menstrual blood-specific markers (MMP7, MMP10, STC1) remained detectable in mixtures up to a 50:1 ratio of peripheral blood to menstrual blood. Based on these findings, we established the following criteria: samples were classified as menstrual blood when both housekeeping genes were stably detected, together with at least three peripheral blood-specific markers and at least three menstrual-specific markers were co-detected. Overall, a universal criterion for distinguishing menstrual blood from peripheral blood samples was established through cross-platform validation in this study. Our findings offer a critical reference for the cross-platform adaptation and standardization of RNA-based menstrual blood identification in forensic applications.

Authors

Niu Gao (School of Forensic Medicine, Shanxi Medical University, China)
Jinding Liu (School of Forensic Medicine, Shanxi Medical University, China)
Jiangwei Yan (School of Forensic Medicine, Shanxi Medical University, China)

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P245 – Cross‑platform Validation of a mRNA Assay for Differentiating Menstrual Blood from Peripheral Blood

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