Impact of nucleic acid extraction platforms on hepatitis virus genome detection

J Virol Methods. 2019 Nov:273:113715. doi: 10.1016/j.jviromet.2019.113715. Epub 2019 Aug 13.

Abstract

Detection and quantification of viral nucleic acids are important for diagnosing current viral infections and monitoring response to antiviral therapy. Automated nucleic acid extraction and purification platforms are routinely used during the first step in these processes in clinical and research laboratories. Here, we compare the extraction efficiencies of four MagNA Pure magnetic bead-based nucleic acid extraction platforms and associated kits using samples positive for nucleic acids from HAV, HBV, HCV, HDV, and HEV. These five hepatitis viruses are diverse in their virion structures and type of nucleic acid that compose their genomes. We found that the most efficient nucleic acid extraction platform and corresponding kit, when averaged across all tested viruses, was the MagNA Pure 96, which yielded twice as much detectable nucleic acid as the other platforms. However, the relative efficiencies of the different platforms varied by virus type, suggesting that an extraction platform that is more efficient for one virus type will not necessarily function better with a different virus type. Our results show that the choice of a nucleic acid extraction platform influences the sensitivity of the methodology and has the potential to generate false-negative results especially in samples with low levels of viral nucleic acids.

Keywords: Nucleic acid testing; Viral hepatitis.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA, Viral / isolation & purification
  • Genome, Viral*
  • Hepatitis Viruses / isolation & purification*
  • Hepatitis, Viral, Human
  • Humans
  • Nucleic Acids / blood
  • Nucleic Acids / isolation & purification*
  • RNA, Viral / isolation & purification
  • Reagent Kits, Diagnostic / standards*
  • Sensitivity and Specificity

Substances

  • DNA, Viral
  • Nucleic Acids
  • RNA, Viral
  • Reagent Kits, Diagnostic