Abstract
To evaluate the identification method using the microRNA markers miR10b and miR135b to distinguish semen stains from menstrual blood, peripheral blood, vaginal fluid and so on body fluid stains. The expression levels of miR10b and miR35b in semen stains and menstrual blood and so on were detected utilizing a real-time quantitative PCR technique with a specific fluorescence-labeled TaqMan probe. RNU6b was used as the internal reference gene; the difference in their expression was analyzed, and the specificity, sensitivity, and detection capability of the techniques were evaluated. The expression of miR10b and miR135b in semen stains was significantly higher than that of other body fluid stains, with a mean value of ΔCт from-6 to-7. However, it ranged from-2 to-4 for other body fluid stains. The initial criteria for judging which semen stains can be identified were determined by analyzing the research results. When the threshold value was set to 0.04, the CT value could be detected in the target genes miR10b, miR135b and in the internal reference gene RNU6b, and CT values are<40, ΔCT[10b-U6]<-5.5, and ΔCT[135b-U6]<-6, respectively, and the semen stain could be identified. The expression levels of miR10b and miR135b are higher in semen with strong tissue specificity; thus, they can be used to differentiate semen stains from other body fluid stains in forensic science.
Highlights
MicroRNAs are a type of endogenous non-coding RNA that has a regulatory function in eukaryotes
When the threshold value was set to 0.04, the CT value could be detected in the target genes miR10b, miR135b and in the internal reference gene RNU6b, and CT values are
MiRNA fragments are short and degrade relatively slowly [5]. Their distributions have the tissue specificity; as such, their unique molecular characteristics may be used as a new way of identifying body fluid stains in forensics
Summary
MicroRNAs (miRNAs) are a type of endogenous non-coding RNA that has a regulatory function in eukaryotes. The length of miRNAs molecule is approximately 20 to 25 nucleotides. They are generated by Dicer enzyme shearing single-stranded RNA precursors, which contain approximately 70 to 90 nucleotides with a hairpin structure [1, 2]. The miRNAs participate in a variety of regulatory pathways, including growth, virus defense, haematopoiesis, organogenesis, cell proliferation, apoptosis, and fat metabolism, and so on. They have been shown to play an important role in tumorigenesis and other related fields.
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