Confirmatory Semen Detection Using Real-Time PCR Technique

Abstract

Objective: To study the specificity of real-time PCR technique for confirmatory semen detection
Methods: Eighteen samples each of blood, saliva, and semen were collected from 18 male volunteers. All 54 samples were subjected to forensic DNA typing and mRNA analysis. DNA templates were amplified using the GlobalFiler® PCR amplification kit. For mRNA analysis protamine 2 (PRM2), a sperm specific gene, was analyzed using β-2-microglobulin gene (B2M), a cellular housekeeping gene, as the internal control.
Results: DNA profiles of all blood, saliva and semen samples from the same volunteers demonstrated exactly similar profiles. For mRNA analysis, B2M was expressed in all 54 samples but only the 18 semen samples showed specific amplification with PRM2.
Conclusions: PRM2 gene was a very effective marker for semen identification. Real-time PCR technique could be suitable for confirmatory detection of semen in medico-legal trace evidence collection.