ABSTRACT
Body fluids analysis is performed in forensic cases to understand the circumstances of the case. Traditional methods of body fluid analysis include colorimetric, enzymatic, immunochromatographic, enzyme-linked immunosorbent assay, and microscopy assays. Next generation sequencing (NGS) methods including pyrosequencing and massively parallel sequencing (MPS) have been developed for multiplex body fluid testing. In addition to methylated DNA targets, RNA targets have also been examined for forensic body fluid analysis. Massively parallel sequencing using the MiSeq and Ion Torrent instruments also offers new opportunities for body fluid analysis. Just as polymerase chain reaction primers have been designed, tested, and multiplexed for human identification and phenotyping applications, they can be designed to target loci such as single nucleotide polymorphisms with demonstrated variations in body fluids. Alternatively, extracted small nuclear RNA or microRNA could be reverse transcribed to cDNA and the targets could be sequenced using MPS using sequencing as is being performed with messenger RNA.
