Life Sciences Commons^™

The Utilization Of Eukaryotic Cell Types In Microbiomes For Semen, Vaginal Fluid, And Menstrual Blood Identification Using 18s Rdna, Ines Benaissa

Master of Science in Forensic Science Directed Research Projects

The accurate identification of body fluids is vital for crime scene investigation and forensic analysis. While many body fluids can be presumptively identified, there is a lack of confirmatory testing for verification. Therefore, the purpose of this study was to determine if body fluids (menstrual blood, vaginal fluid, and semen) could be differentiated based on their eukaryotic microbial communities. The target area of interest was the V9 hyper-variable region of 18S ribosomal DNA (18S rDNA). The samples were extracted, barcoded using a duel-index strategy, and PCR amplified before undergoing high throughput sequencing using the Illumina Miseq FGx instrument. The samples …

A Eukaryotic Signature Based Method For Identification Of Saliva, Feces, And Urine, Alyssa Daniels

Master of Science in Forensic Science Directed Research Projects

The conception of the Human Microbiome Project advanced the understanding of microbial communities in the human body and previous research has established that unique microbial signatures can help distinguish each body fluid. While these signatures have been developed for the prokaryotic microbiome, the next step is the examination of the eukaryotic microbiome. Eukaryotic signatures could provide a greater specificity and statistical weight when discerning between body fluids. These microbial markers can be implemented to develop a confirmatory assay for body fluid identification that works in tandem with other DNA based methods in the forensic workflow. Using a VCU approved IRB …

The Truth About Metagenomics: Quantifying And Counteracting Bias In 16s Rrna Studies, J. Paul Brooks, David J. Edwards, Michael D. Harwich, Maria C. Rivera, Jennifer M. Fettweis, Myrna G. Serrano, Robert Reris, Nihar U. Sheth, Bernice Huang, Phillipe Girerd, Vaginal Microbiome Consortium, Jerome F. Strauss Iii, Kimberly K. Jefferson, Gregory A. Buck

Study of Biological Complexity Publications

Background

Characterizing microbial communities via next-generation sequencing is subject to a number of pitfalls involving sample processing. The observed community composition can be a severe distortion of the quantities of bacteria actually present in the microbiome, hampering analysis and threatening the validity of conclusions from metagenomic studies. We introduce an experimental protocol using mock communities for quantifying and characterizing bias introduced in the sample processing pipeline. We used 80 bacterial mock communities comprised of prescribed proportions of cells from seven vaginally-relevant bacterial strains to assess the bias introduced in the sample processing pipeline. We created two additional sets of 80 …