Simultaneous analysis of micro-RNA and DNA for determining the body fluid origin of DNA profiles

J Forensic Sci. 2013 Jul;58(4):967-71. doi: 10.1111/1556-4029.12160. Epub 2013 May 17.

Abstract

Micro-RNAs (miRNAs) can be specifically expressed in forensically relevant body fluids such as blood or saliva. The aim of the study was to develop a simultaneous extraction and analysis protocol that allows for the acquisition of a DNA profile and the identity of the body fluid using a single process. DNA and micro-RNA were extracted from blood and saliva before undergoing a cDNA synthesis step by using stem-loop reverse transcription PCR. The resulting extracts containing DNA and cDNA synthesized from body fluid-specific miRNA markers then underwent standard STR analysis using a modified ABI AmpFℓSTR(®) NGM SElect™ kit. In all samples, a full DNA profile was obtained along with additional peaks corresponding to the miRNA marker targeted. In all cases, blood samples profiled exhibited a peak indicating the presence of the blood-specific miRNA marker and the saliva sample profiled exhibited a peak indicating the presence of the saliva-specific miRNA marker.

Keywords: DNA; body fluid identification; co-isolation; forensic genetics; forensic science; micro-RNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blood Chemical Analysis*
  • DNA / isolation & purification*
  • DNA Fingerprinting / methods*
  • Electrophoresis, Capillary
  • Humans
  • MicroRNAs / isolation & purification*
  • Microsatellite Repeats
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcription
  • Saliva / chemistry*

Substances

  • MicroRNAs
  • DNA