Background: The discovery of forensic DNA typing evolved molecular biology far beyond
what could have been expected in terms of its forensic application, and now there exists other developments
in molecular biology which are ready for application to forensic challenges. One such challenge
is the identification of the body fluid source of stains recovered from evidence items and crime
scenes. Currently, there are significant efforts in the research field to develop novel methods for the
molecular identification of body fluids, with microRNAs (miRNAs) revealing great potential.
MiRNAs have been shown to have high tissue specificity and are less susceptible to degradation as a
result of their small size, which infers great advantages to their potential role for identifying forensically
relevant body fluids.
Objective: This study investigated the isolation and amplification of miRNAs from forensically relevant
Method: Venous blood, menstrual blood, semen, saliva, and vaginal material samples were extracted
using; miRNeasy® mini kit (Qiagen), mirVana™ miRNA isolation kit (Ambion), and a modified mir-
Vana™ method, and the quality/quantity of isolated miRNA was determined. miRNAs previously
identified to show specificity for particular forensically relevant body fluids were examined. Real
Time-Quantitative PCR (RT-qPCR) was performed targeting 5 miRNAs of interest, miR-451, miR-
412, miR-891a, miR-205 and miR-124a.
Results: This study identified the miRNeasy® mini kit as the optimal method of the three methods investigated
for the extraction of miRNAs from body fluids and further validates a selection of miRNAs
previously suggested as potential biomarkers.
Conclusion: This research highlights the potential of miRNAs as novel markers for the identification
of forensically relevant body fluids.