Identification of Suitable Reference Gene for Normalization of microRNAs in Forensically Revenant Body Fluids by Reverse Transcription-Quantitative Polymerase Chain Reaction

Abstract

Body fluids are well known to be a very useful evidence type in criminal investigations.MicroRNAs are small non-coding RNAs that have previously been identified as possible markers for forensically related body fluid detection.Adequate data normalization is essential to reduce nonbiological technical changes to perform reliable miRNA expression analyzes using a quantitative PCR.The best reference genes are those that are biologically stable and whose expression does not vary across samples. The current study investigated ten selected candidate reference genes that could be used to normalize miRNA expression using an RT qPCR assay, including: SNORD-48; SNORD-44; SNORD-47; U6-2; hsa-miR-26b-5p; hsa-miR-92a-3p; hsa-miR-93-3p; hsa-miR-191-5p; hsa-miR-21; hsa-miR-484.The present results revealed that the cycle threshold (Ct) values of most genes ranged from 3.12 to 27.03, and identify SNORD-47 as a suitable reference gene for all sample types included venous blood, semen, and vaginal secretion.