- •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.
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.
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- Body fluid identification in forensics.BMB Rep. 2012; 45: 545-553
- A stepwise strategy to distinguish menstrual blood from peripheral blood by Fisher’s discriminant function.Int. J. Leg. Med. 2020; 134: 845-851
- Screening and confirmation of microRNA markers for distinguishing between menstrual and peripheral blood.Forensic Sci. Int. Genet. 2017; 30: 24-33
- Potential applications of microRNA profiling to forensic investigations.RNA. 2020; 26: 1-9
- Analysis of body fluids for forensic purposes: from laboratory testing to non-destructive rapid confirmatory identification at a crime scene.Forensic Sci. Int. 2009; 188: 1-17
- Molecular approaches for forensic cell type identification: on mRNA, miRNA, DNA methylation and microbial markers.Forensic Sci. Int. Genet. 2015; 18: 21-32
- MicroRNAs: small RNAs with a big role in gene regulation.Nat. Rev. Genet. 2004; 5: 522-531
- Screening of exosomal microRNAs from colorectal cancer cells.Cancer Biomark. 2016; 4: 427-435
- Micro-RNA – a potential for forensic science?.Forensic Sci. Int. 2010; 203: 106-111
- Circulating microRNA for the identification of forensically relevant body fluids.Methods Mol. Biol. 2013; 1024: 221-234
- Forensic miRNA: potential biomarker for body fluids?.Forensic Sci. Int. Genet. 2015; 14: 1-10
- Identification of forensically relevant body fluids using a panel of differentially expressed microRNAs.Anal. Biochem. 2009; 387: 303-314
- MicroRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation.Int. J. Leg. Med. 2010; 124: 217-226
- Screening and confirmation of microRNA markers for forensic body fluid identification.Forensic Sci. Int. Genet. 2013; 7: 116-123
- Differentiation of five body fluids from forensic samples by expression analysis of four microRNAs using quantitative PCR.Forensic Sci. Int. Genet. 2016; 22: 89-99
- Investigating the isolation and amplification of microRNAs for forensic body fluid identification.Microrna. 2018; 7: 187-194
- The identification of menstrual blood in forensic samples by logistic regression modeling of miRNA expression.Electrophoresis. 2014; 35: 3087-3095
- MiR-21-5p and miR-126a-3p levels in plasma and circulating angiogenic cells: relationship with type 2 diabetes complications.Oncotarget. 2015; 6: 35372-35382
- Bone morphogenetic protein 4 promotes vascular smooth muscle contractility by activating microRNA-21 (miR-21), which down-regulates expression of family of dedicator of cytokinesis (DOCK) proteins.J. Biol. Chem. 2012; 287: 3976-3986
- Comparative evaluation of different extraction and quantification methods for forensic RNA analysis.Forensic Sci. Int. Genet. 2015; 16: 195-202
- Predicting effective microRNA target sites in mammalian mRNAs.eLife. 2015; 4e05005
- miRDB: an online resource for microRNA target prediction and functional annotations.Nucleic Acids Res. 2015; 43: D146-D152
- miRTarBase: a database curates experimentally validated microRNA-target interactions.Nucleic Acids Res. 2011; 39: D163-D169
- Cytoscape: a software environment for integrated models of biomolecular interaction networks.Genome Res. 2003; 13: 2498-2504
- The ratio of miR-21/miR-24 as a promising diagnostic and poor prognosis biomarker in colorectal cancer.Eur. Rev. Med. Pharmacol. Sci. 2018; 22: 8649-8656
- MicroRNA expression ratio is predictive of head and neck squamous cell carcinoma.Clin. Cancer Res. 2009; 15: 2850-2855
- MicroRNA ratios distinguish melanomas from nevi.J. Investig. Dermatol. 2020; 140: 164-173
- Associations of circulating plasma microRNAs with age, body mass index and sex in a population-based study.BMC Med. Genom. 2015; 8: 61
- Age estimation using bloodstain miRNAs based on massive parallel sequencing and machine learning: a pilot study.Forensic Sci. Int. Genet. 2020; 47102300
- Age‐associated microRNA expression in human peripheral blood is associated with all‐cause mortality and age‐related traits.Aging Cell. 2018; 17e12687
- Characterising the fluctuation of microRNA expression throughout a full menstrual cycle.Forensic Sci. Int. Genet. 2015; Suppl. 5: Se264-Se266
- Massively parallel sequencing of microRNA in bloodstains and evaluation of environmental influences on miRNA candidates using real-time polymerase chain reaction.Forensic Sci. Int. Genet. 2019; 38: 32-38
- The stability and persistence of blood and semen mRNA and miRNA targets for body fluid identification in environmentally challenged and laundered samples.Leg. Med. 2019; 38: 45-50
- MRNA heptaplex protocol for distinguishing between menstrual and peripheral blood.Forensic Sci. Int. Genet. 2014; 13: 53-60
- A human protein atlas for normal and cancer tissues based on antibody proteomics.Mol. Cell Proteom. 2005; 27: 1920-1932
- Database resources of the National Center for Biotechnology Information.Nucleic Acids Res. 2010; 38: D5-D16
Published online: December 13, 2021
Accepted: December 10, 2021
Received in revised form: November 12, 2021
Received: June 30, 2021
© 2021 Published by Elsevier B.V.