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. 2023 Aug 7;45(8):6526-6537.
doi: 10.3390/cimb45080411.

An mRNA Profiling Study of Vaginal Swabs from Pre- and Postmenopausal Women

Elena Chierto  1 , Federica Alessandrini  2 , Carla Bini  3 , Eugenia Carnevali  4 , Matteo Fabbri  5 , Paolo Fattorini  6 , Pierangela Grignani  7 , Francesca Scarnicci  8 , Pamela Tozzo  9 , Andrea Verzeletti  10 , Susi Pelotti  3 , Loredana Buscemi  11 , Carlo Robino  1
Affiliations

An mRNA Profiling Study of Vaginal Swabs from Pre- and Postmenopausal Women

Elena Chierto et al. Curr Issues Mol Biol. .

Abstract

Body fluid identification by means of mRNA profiling provides valuable supplementary information in forensic investigations. In particular, the detection of vaginal mucosa mRNA markers is highly relevant in sexual assault cases. Although the vagina undergoes characteristic age-related physiological changes over a lifetime, few studies have evaluated the efficacy of vaginal mRNA markers in women of different ages. In this multicentric study, a 19-plex mRNA profiling assay including vaginal-specific markers (CYP2B7P1, MUC4, MYOZ1) was tested in a collection of 6-20-month-old vaginal swabs obtained from pre- (n = 84) and postmenopausal (n = 55) female volunteer donors. Overall, participating laboratories were able to correctly identify ~85% of samples as vaginal mucosa by mRNA profiling. The assay's success rate did not differ between the two age groups and was not affected by the time interval between swab collection and RNA analysis. MYOZ1 resulted a less sensitive vaginal marker compared to MUC4 and CYP2B7P1. A significant relative increase in the contribution to the total amplification signal was observed for MUC4, compared to CYP2B7P1 and MYOZ1, in postmenopausal women. Observation of other body fluids and tissues different from vaginal mucosa was also evaluated in connection to information on previous sexual activity and menstrual cycle phase at the time of sampling.

Keywords: CYP2B7P1; MUC4; MYOZ1; body fluid identification; mRNA profiling; vaginal mucosa.

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Conflict of interest statement

The authors have declared no conflict of interest.

Figures

Figure 1
Figure 1
mRNA profiling success rates (vaginal mucosa observed, according to the scoring method) across laboratories in the whole sample (Total) and within subcategories of PR-M and PO-M women. Data are expressed as mean ± SE.
Figure 2
Figure 2
Distribution of tested samples according to time interval between swab collection and RNA extraction expressed in months (x axis) and mRNA profiling outcome considering the total number of samples (a), PR-M samples (b) and PO-M samples (c). Vaginal mucosa was “observed” (black) or “not observed” (gray), according to the scoring method.
Figure 3
Figure 3
Percentage of peaks above the analytical threshold for the vaginal markers CYP2B7P1 (a), MUC4 (b), and MYOZ1 (c) in mRNA profiling replicates of PR-M (n = 84) and PO-M (n = 55) vaginal samples. Relative contribution of each gene to total peak height for the vaginal markers CYP2B7P1 (d), MUC4 (e), and MYOZ1 (f) in mRNA profiling replicates of PR-M (n = 75) and PO-M (n = 50) vaginal samples. Data are expressed as mean ± SE. *: p < 0.05 (t-test).
Figure 4
Figure 4
Percentage of peaks above the analytical threshold for the vaginal markers CYP2B7P1, MUC4, and MYOZ1 in PR-M (a) and PO-M vaginal samples (b). Relative contribution of each gene to total peak height for the vaginal markers CYP2B7P1, MUC4, and MYOZ1 in PR-M (c) and PO-M vaginal samples (d). Data are expressed as mean ± SE. *: p < 0.05; **: p < 0.01; ***: p < 0.001; (a) ANOVA with Tukey post hoc test; (b–d) Kruskal–Wallis with Dunn’s post hoc test).
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