mRNA MPS Tissue Identification Assay to Aid in the Investigation of Traumatic Injuries

Abstract Molecular analysis of the RNA transcriptome from a putative tissue fragment should permit assignment to a specific organ since each tissue will exhibit a unique pattern of gene expression. Determination of the organ source of tissues from crime scenesmay aid in shooting and stabbinginvestigations. We have developed a new prototype massively parallel sequencing(MPS) mRNA profiling assay for organ tissue identification, designed to definitively identify 13 organ/ tissue typesusing a targeted panel of 48 mRNA biomarkers. The identifiable organs and tissues include brain, spinal cord, lung, trachea, liver, skeletal muscle, heart, kidney, adipose, intestine, stomach, skin and spleen. The biomarkers were chosen after iterative specificity testing of numerous candidate genes in various tissue types. The assay is very specific with little cross reactivity with non-targeted tissue, and can detect RNA mixtures from different tissues, including two- to five-tissue admixtures. The sensitivity of the assay was evaluated as well as assay reproducibility between library preparations and sequencing runs. We also demonstrate the ability of the assay to successfully identify the tissue source of origin in cadaver samples, tissue samples with varying post mortem intervals(PMI) and mock and bona fidecasework samples. We are using the data to train a multivariate statistical model that predicts the tissue typebased on the mRNA profile. By considering co-expression of markers the model can recognize distinct expression patterns in each tissue.