The Human Virome as Trace Evidence in Forensic Investigation

As submitted by the proposer: This proposal is designed to address the ongoing need to create forensically relevant linkages between persons, places, and objects by developing the untapped potential of the human viral microbiome (virome). The human virome is a source of rich genetic diversity that has not been examined to determine if it is stable, transferable, and provides sufficient power of discrimination to be used as an alternative to traditional human forensic deoxyribonucleic acid (DNA) tests when conditions for such tests are not suitable. There is a complete lack of empirical data on the human virome with regard to its suitability to forensic applications. The proposed work will address this gap in our knowledge by generating data in the previously mentioned areas of stability, transfer, and discrimination. The proposal hypothesizes that the genetic diversity contained in each human being’s particular skin virome can be translated into a pattern profile that can be used to discriminate them from all others and that the virome profile can be detected when standard human DNA samples are not viable. The work will be done with protocols, instrumentation, and analysis methods that are either already in forensic DNA laboratories or can be readily assimilated. This project will recruit 60 adult human subjects, 30 female and 30 male, and collect skin virome samples and transfer skin virome samples from three different sites at five time points over a period of six months. These samples will be processed for viral DNA extraction and subsequent next generation sequencing. The researchers will use a robust two-pronged approach to viral sequence pre-processing and determination of virome composition and abundances. Statistical evaluation and testing will be used to assess the stability, transfer, and discriminatory power of the virome adjusted for subject characteristics and time of sampling. Based on data and results, the researchers will create and host a web-accessible public database of human skin virome profiles for the estimation of composition and rarity of such profiles. The researchers anticipate scholarly publications, presentations, and access to data processing and analysis pipelines. This project will further our understanding of the human viral microbiome, address the critical barrier of individual identification when conditions for human DNA testing are poor, and develop new technology for the use of the human virome as a tool for forensic applications. Note: This project contains a research and/or development component, as defined in applicable law, and complies with Part 200 Uniform Requirements - 2 CFR 200.210(a)(14). ca/ncf