Highlights
- •A cSNPs typing system based on vaginal secretion (VS) specific mRNA markers has been established for detecting the VS.
- •This assay has favorable specificity and sensitivity in VS-containing mixtures deconvolution.
- •This assay provides a convenient method for researchers in their identification of VS.
- •The polymorphicmarkers in the body-fluid specific mRNA could be used to identify the donorinvolved in the mixtures.
Abstract
In recent years, RNA profiling has become an important technique in identifying the
origin of human body fluids/tissues. Both perpetrators and victims can be identified
from stains involving vaginal secretions (VS), such as sperm/VS mixtures left on condoms,
bed sheets, or papers, etc. Body fluid specific RNA typing could link the source of
body fluids/tissues and the identity of the donor. In this study, we aimed to trace
the donor of VS in mixture stains using body fluid-specific mRNA markers and construct
a coding single nucleotide polymorphism (cSNP) typing system for VS. We screened 8
VS-specific mRNA biomarkers (MUC4, SFTA2, CYP2A6, MYOZ1, FUT6, ESR1, SPINK5, and SERPINB13)
encompassing 18 cSNPs. The RNA obtained from various body fluid/tissue samples was
treated with reverse transcription polymerase chain reaction (RT-PCR) and then followed
by a multiplex PCR and SNaPshot mini-sequencing assay. The detection limit of the
assay was 0.08 ng RNA. For single-source body fluid, the positive cSNP typing was
only shown in VS and void in non-VS body fluids/tissues. For laboratory-generated
VS-containing mixtures, the minor VS contributor could be successfully detected at
a ratio of 1:10–1:500. We also confirmed the concordance of DNA typing and mRNA typing
for the cSNPs in this system. In summary, we established an 18-cSNP typing system
for VS with high sensitivity and specificity, which could identify both the donor
and the tissue origin simultaneously. This was shown to be a powerful tool for identifying
the VS donor in those VS-containing mixture stains.
Abbreviations:
VS (vaginal secretion), CE (capillary electrophoresis), NGS (next-generation sequencing), EPG (electrophoretogram), RFU (relative fluorescence units)Keywords
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References
- Forensic molecular biomarkers for mixture analysis.Forensic Sci. Int Genet. 2019; 41: 107-119https://doi.org/10.1016/j.fsigen.2019.04.003
- Chapter 2—a deep analysis of the basic causes of interpretation errors.in: Gill P. Misleading DNA Evidence. Academic Press, San Diego2014
- Biosensor developments: application to prostate-specific antigen detection.TRENDS Biotechnol. 2007; 25: 125-131https://doi.org/10.1016/j.tibtech.2007.01.004
- A multiplex (m)RNA-profiling system for the forensic identification of body fluids and contact traces.Forensic Sci. Int Genet. 2012; 6: 565-577https://doi.org/10.1016/j.fsigen.2012.01.009
- Highly specific mRNA biomarkers for the identification of vaginal secretions in sexual assault investigations.Sci. JUSTICE. 2013; 53: 14-22https://doi.org/10.1016/j.scijus.2012.03.007
- mRNA profiling using a minimum of five mRNA markers per body fluid and a novel scoring method for body fluid identification.Int. J. Leg. Med. 2013; 127: 707-721https://doi.org/10.1007/s00414-012-0794-3
- Messenger RNA biomarker signatures for forensic body fluid identification revealed by targeted RNA sequencing.Forensic Sci. Int.: Genet. 2018; 34: 206-221https://doi.org/10.1016/j.fsigen.2018.02.020
- An evidence based strategy for normalization of quantitative PCR data from miRNA expression analysis in forensic organ tissue identification.Forensic Sci. Int Genet. 2014; 13: 217-223https://doi.org/10.1016/j.fsigen.2014.08.005
- Characterization of tissue-specific biomarkers with the expression of circRNAs in forensically relevant body fluids.Int. J. Leg. Med. 2019; 133: 1321-1331https://doi.org/10.1007/s00414-019-02027-y
- A novel multiplex assay system based on 10 methylation markers for forensic identification of body fluids.J. FORENSIC Sci. 2021; https://doi.org/10.1111/1556-4029.14872
- Microbiome-based body fluid identification of samples exposed to indoor conditions.Forensic Sci. Int Genet. 2019; 40: 105-113https://doi.org/10.1016/j.fsigen.2019.02.010
- RNA/DNA co-analysis from human menstrual blood and vaginal secretion stains: results of a fourth and fifth collaborative EDNAP exercise.Forensic Sci. Int Genet. 2014; 8: 203-212https://doi.org/10.1016/j.fsigen.2013.09.009
- Evaluation of mRNA marker specificity for the identification of five human body fluids by capillary electrophoresis.Forensic Sci. Int.: Genet. 2012; 6: 452-460https://doi.org/10.1016/j.fsigen.2011.09.007
- Identification of coding region SNPs from specific and sensitive mRNA biomarkers for the deconvolution of the semen donor in a body fluid mixture.Forensic Sci. Int Genet. 2021; 52102483https://doi.org/10.1016/j.fsigen.2021.102483
- A method of identifying the blood contributor in mixture stains through detecting blood-specific mRNA polymorphism.electrophoresis. 2020; 41: 1364-1373https://doi.org/10.1002/elps.202000053
- Current Methods for Body Fluid Identification Related to Sexual Crime: Focusing on Saliva, Semen, and Vaginal Fluid.Diagnostics. 2020; 10: 693https://doi.org/10.3390/diagnostics10090693
- Recovery and stability of RNA in vaginal swabs and blood, semen, and saliva stains.J. FORENSIC Sci. 2008; 53: 296-305https://doi.org/10.1111/j.1556-4029.2007.00652.x
- Multiplex mRNA profiling for the identification of body fluids.FORENSIC Sci. Int. 2005; 152: 1-12https://doi.org/10.1016/j.forsciint.2005.02.020
- RNA Profiling for the Identification of the Tissue Origin of Dried Stains in Forensic Biology.Forensic Sci. Rev. 2010; 22: 145-157
- mRNA profiling of mock casework samples: Results of a FoRNAP collaborative exercise.Forensic Sci. Int Genet. 2021; 50102409https://doi.org/10.1016/j.fsigen.2020.102409
- Genome-wide mRNA profiling and multiplex quantitative RT-PCR for forensic body fluid identification.Forensic Sci. Int.: Genet. 2013; 7: 143-150https://doi.org/10.1016/j.fsigen.2012.09.001
- Comparing self-directed methods for training staff to create graphs using Graphpad Prism.J. Appl. Behav. Anal. 2019; 52: 188-204https://doi.org/10.1002/jaba.522
- Haploview: Visualization and analysis of SNP genotype data.Cold Spring Harb. Protoc. 2009; 2009: p71https://doi.org/10.1101/pdb.ip71
- [Genetic Parameters of SNP Loci in Next Generation Sequencing Kits and Their Comparison].Fa Yi Xue Za Zhi. 2018; 34: 242-247https://doi.org/10.12116/j.issn.1004-5619.2018.03.005
- Body fluid identification in forensics.BMB Rep. 2012; 45: 545-553https://doi.org/10.5483/bmbrep.2012.45.10.206
- Assigning forensic body fluids to donors in mixed body fluids by targeted RNA/DNA deep sequencing of coding region SNPs.Int. J. Leg. Med. 2020; 134: 473-485https://doi.org/10.1007/s00414-020-02252-w
- Development of highly sensitive and specific mRNA multiplex system (XCYR1) for forensic human body fluids and tissues identification.PLoS One. 2014; 9e100123https://doi.org/10.1371/journal.pone.0100123
- How specific are the vaginal secretion mRNA-markers HBD1 and MUC4?.Forensic Sci. Int.: Genet. Suppl. Ser. 2009; 2
Article info
Publication history
Published online: April 02, 2022
Accepted:
March 28,
2022
Received in revised form:
March 27,
2022
Received:
October 15,
2021
Identification
Copyright
© 2022 Elsevier B.V. All rights reserved.
