Highlights
- •The copy number ratio of miR-451a to miR-21–5p was first used as the reference standard for human blood sample identification.
- •The identification accuracy of the new strategy was almost 100%.
- •The identification accuracy of the new strategy was rarely affected by physiological and environmental factors.
- •The blood sample included 0.2 ng RNA was enough for this new strategy.
Abstract
Distinction between menstrual blood and peripheral blood is vital for forensic casework,
as it could provide strong evidence to figure out the nature of some criminal cases.
However, to date no single blood-specific gene, including the most variable microRNAs
(miRNAs) could work well in identification of blood source. In this study, we developed
a new strategy for identification of human blood samples by using the copy number
ratios of miR-451a to miR-21–5p based on 133 samples, including 56 menstrual blood
and 47 peripheral blood, as well as 30 non-blood samples of saliva (10), semen (10)
and vaginal secretion (10). The cut-off value and efficacy of the identification strategy
were determined through receiver operating characteristic (ROC) analysis. Our results
showed that when the miR-451a/miR-21–5p ratio below 0.929, the sample should be non-blood.
In contrast, when the miR-451a/miR-21–5p ratio above 0.929 and below 10.201, the sample
should be menstrual blood; and when this ratio above 10.201, the sample should be
peripheral blood. External validation using 86 samples (62 menstrual blood and 24
peripheral blood samples) fully supported this strategy with the 100% sensitivity
and 100% specificity. We confirmed that this result accuracy was not affected by various
potential confounding factors of samples and different experimental platforms. We
showed that 0.2 ng of total RNA from menstrual blood and peripheral blood was sufficient
for qPCR quantification. In conclusion, our results provide an accurate reference
to distinguish menstrual blood from peripheral blood for forensic authentication.
Keywords
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Article info
Publication history
Published online: December 13, 2021
Accepted:
December 10,
2021
Received in revised form:
November 12,
2021
Received:
June 30,
2021
Identification
Copyright
© 2021 Published by Elsevier B.V.
