Medical, Molecular and Forensic Sciences, Murdoch University, 90 South Street, Murdoch 6150, Australia.
Australian National Phenome Centre and Computational and Systems Medicine, Health Futures Institute, Harry Perkins Institute of Medical Research, Perth, Western Australia 6150, Australia.
Forensic Sci Int. 2023 Jul;348:111711. doi: 10.1016/j.forsciint.2023.111711. Epub 2023 Apr 27.
The detection and recovery of male DNA post-assault is important in sexual assault investigations, particularly where an offender is unknown to the victim. The collection of DNA evidence often occurs when the female victim undergoes a forensic medical assessment. Analysis regularly results in mixed autosomal DNA profiles with both victim and perpetrator DNA, often making it difficult to interpret a male profile suitable for DNA database searching. While short tandem repeat (STR) profiling of the male Y-chromosome is often used to overcome this challenge, successful identification of an individual can be hindered by the paternal inheritance pattern of Y-STRs and small Y-STR databases. Human microbiome research has suggested that a person's microbial diversity is unique. Therefore microbiome analysis using Massively Parallel Sequencing (MPS) could serve as a useful adjunct method of perpetrator identification. This study aimed to identify bacteria taxa that were unique to each participant and compare the bacterial communities found on their genitals both pre- and post-coitus. Samples were collected from six male-female sexual partner pairs. Volunteers were asked to self-collect low vaginal (females) and penis shaft and glans (males) samples before and after intercourse. Samples were extracted using the PureLink™ Microbiome DNA Purification Kit. Extracted DNA underwent library preparation using primers targeting the V3-V4 hypervariable regions of the bacterial 16S rRNA gene (∼450 bp). Libraries were sequenced on the Illumina MiSeq® platform. From the sequence data derived, statistical analysis was performed to investigate if bacteria sequences could be used to infer contact between each male-female pairing. Unique bacterial signatures were detected in low frequencies (<1%) in male and female participants pre-coitus. The data indicated a significant disruption to microbial diversity post-coitus in all samples. A transfer of the female microbiome during intercourse was most significant. As expected, one couple who did not use a barrier contraceptive yielded the most microbial transfer and disruption to diversity demonstrating a proof-of-concept in the utility of microbiome interrogation for sexual assault cases. Further genomic analysis is needed to confirm species and subspecies classification of bacteria that may produce a unique microbial profile that could then be used to identify a specific individual.
在性侵犯调查中,检测和恢复男性 DNA 非常重要,特别是在受害者不认识犯罪者的情况下。通常在女性受害者接受法医评估时收集 DNA 证据。分析结果通常是混合的常染色体 DNA 图谱,既有受害者的 DNA,也有犯罪者的 DNA,这使得解释适合 DNA 数据库搜索的男性图谱变得非常困难。虽然短串联重复序列(STR)分析男性 Y 染色体通常用于克服这一挑战,但 Y-STR 的父系遗传模式和较小的 Y-STR 数据库会阻碍对个体的成功识别。人类微生物组研究表明,一个人的微生物多样性是独特的。因此,使用大规模平行测序(MPS)进行微生物组分析可以作为一种有用的犯罪者识别辅助方法。本研究旨在确定每个参与者特有的细菌分类群,并比较他们的生殖器在性交前后的细菌群落。从六对男女性伴侣中采集样本。志愿者被要求在性交前后自行采集阴道(女性)和阴茎轴和龟头(男性)样本。使用 PureLink™微生物 DNA 纯化试剂盒提取样品。使用针对细菌 16S rRNA 基因(~450 bp)的 V3-V4 高变区的引物制备提取 DNA 的文库。文库在 Illumina MiSeq®平台上进行测序。从获得的序列数据中,进行统计分析以调查细菌序列是否可用于推断每个男女配对之间的接触。在性交前,男性和女性参与者的低频率(<1%)检测到独特的细菌特征。数据表明,所有样本的微生物多样性在性交后都受到显著破坏。在性交过程中,女性微生物组发生了转移。正如预期的那样,一对没有使用屏障避孕措施的夫妇产生了最多的微生物转移和多样性破坏,证明了微生物组询问在性侵犯案件中的实用性的概念验证。需要进一步的基因组分析来确认可能产生独特微生物图谱的细菌的种和亚种分类,然后可以使用该图谱来识别特定个体。