Effect of mismatch between types of viral nucleic acid and intended targets of extraction kits on polymerase chain reaction-based testing

Biotechniques. 2022 Aug;73(2):75-79. doi: 10.2144/btn-2022-0006. Epub 2022 Jun 28.

Abstract

Lab personnel generally select an extraction kit based on the nucleic acid (NA) type of the target. This study investigated the effect of mismatch between the NA type of the target and the intended target NA of the extraction kit on the polymerase chain reaction outcome. DNA, RNA and total NA extraction kits manufactured by the same company were used to isolate NA from serial dilutions of four viruses representing different genome types. All extracts were tested for the viruses by either conventional or real-time polymerase chain reactions with and without reverse transcription. While the DNA kit specifically isolated DNA from samples, the RNA kit extracted both DNA and RNA as efficiently as the total NA kit, suggesting that RNA kits can be an economical alternative.

Keywords: bovine herpesvirus type 1; nucleic acid extraction kit; polymerase chain reaction; porcine circovirus type 2; porcine epidemic diarrhea virus; porcine rotavirus type A.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Nucleic Acids* / genetics
  • RNA, Viral / genetics
  • Reagent Kits, Diagnostic
  • Real-Time Polymerase Chain Reaction
  • Sensitivity and Specificity
  • Viruses* / genetics

Substances

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