Date of Submission
Master of Science in Forensic Science
David San Pietro
Body fluids--Analysis, MicroRNA
Body fluid identification is an important aspect of forensic investigations because it assists with the reconstruction of crime scenes and can refute and/or support witness statements. Currently, there is no universal method for body fluid identification. Each body fluid has several tests for its identification, both presumptive and confirmatory. A universal method for body fluid identification that is sensitive, specific, efficient, and minimally destructive is necessary.
In recent years miRNAs have been heralded as novel biomarkers for the identification of body fluids. Several research groups across the US and internationally have been formed to determine which miRNAs are suitable for body fluid identification, however there has been little agreement between them in their conclusions. With the advent of Next Generation Sequencing (NGS), it is now possible to sequence all forensically relevant body fluids for both known and novel miRNAs, with the expressed interest to identify panels of miRNAs for each body fluid.
The goal of this study was to determine if it was feasible to use NGS in the form of the MiSeq FGx platform (Illumina) to determine a panel of miRNAs that are specific to forensically relevant body fluids including venous blood, semen, saliva, vaginal fluid, and menstrual blood. Subsequently, a panel of miRNAs was identified from previously published research that were reported to have potential applications to forensic body fluid identification. Ultimately, it was the goal of this study to determine if the chosen miRNAs were suitable for body fluid identification. This was done using RT-qPCR, which is the preferred method of miRNA expression analysis.
It was determined that the MiSeq FGx was not an optimal NGS platform for this purpose, as no miRNAs were identified. The RT-qPCR validation of the chosen miRNAs resulted in both agreement and disagreement with previously published results, illustrating the need for more extensive research to determine an appropriate panel of miRNAs for body fluid identification.
Johannsen, Karly, "Small RNA Sequencing and RT-qPCR Validation of Forensically Relevant Body Fluids" (2019). Master's Theses. 115.