Highlights
- •Sexual assault with anal penetration is closely related to child or male victims.
- •Few procedures use biological samples to identify anal penetration.
- •The identification of rectal mucosa might help to prove such a sexual assault.
- •Rectal mucosa mRNA markers were screened and a multiplex RT-PCR was developed.
- •Precise determination of rectal mucosa could be performed with this procedure.
Abstract
Sexual assault with anal penetration is closely related to child sexual abuse or male
victims. However, it is difficult to prove such an act by using biological samples
collected from the surface of a suspected object because procedures for identifying
rectal mucosa have not been developed sufficiently. Therefore, for the specific identification
of rectal mucosa, mRNA markers reported to be characteristically expressed in the
rectum were screened and a multiplex RT-PCR procedure was developed for the simultaneous
determination of those candidate markers. The detectability and specificity of rectal
mucosa candidate markers were evaluated using rectal mucosa samples and forensically
relevant body fluids. Diluted or mixed samples were also tested to evaluate the applicability
of this procedure for forensic casework. As a result, simultaneous amplification and
determination of the selected candidates (PHGR1, MUC13, CLCA1, MEP1A, CDX1, and ZG16) and reference gene were successfully performed using a multiplex RT-PCR assay combined
with capillary electrophoresis and fragment analysis. Applying the cutoff values,
none of the other body fluids cross-reacted with rectal mucosa candidate markers.
Because the low sensitivity and detectability of some candidate markers could be compensated
for by their simultaneous detection, all six candidate markers were considered to
be applicable as rectal mucosa markers. Besides, the developed assay should not be
performed on suspicious fecal samples directly because these markers could be positive
in the fecal samples themselves. The developed multiplex RT-PCR procedure might not
be suitable for minute or diluted samples; however, it might be resistant to contamination
with sexual assault-related body fluids. In conclusion, the simultaneous determination
of selected rectal mucosa markers with a biological sample collected from the surface
of a suspected object could be beneficial for criminal investigation of sexual assault
with anal penetration.
Keywords
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Article info
Publication history
Published online: April 15, 2022
Accepted:
April 13,
2022
Received in revised form:
March 28,
2022
Received:
December 16,
2021
Identification
Copyright
© 2022 Elsevier B.V. All rights reserved.
