Use of internal standard RNA molecules for the RT-PCR amplification of the faeces-borne RNA viruses

J Virol Methods. 2002 Jul;104(2):107-15. doi: 10.1016/s0166-0934(02)00016-2.

Abstract

The diagnostic system based on reverse transcription (RT)-PCR has been used widely for the detection of viral genomes of faecal-borne RNA viruses. However, faecal specimens often produce both false positive and false negative results. Therefore, there is a need for a diagnosis procedure that can control for 'false-results'. In this study, an internal standard RNA that can serve as a non-competitive positive template was developed and used directly to detect faecal-borne RNA viruses without noticeable competitive inhibition of the target viral genome. These results suggest that the internal standard RNA is a useful standard molecule when undertaking diagnostic qualitative RT-PCR procedures for enteroviruses and related faecal-borne RNA viruses.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Cloning, Molecular
  • DNA Primers
  • DNA, Viral / genetics
  • Feces / virology*
  • Humans
  • Molecular Sequence Data
  • RNA Virus Infections / diagnosis*
  • RNA Viruses / classification
  • RNA Viruses / genetics
  • RNA Viruses / isolation & purification*
  • RNA, Viral / genetics
  • RNA, Viral / isolation & purification*
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Rotavirus / genetics
  • Rotavirus / isolation & purification
  • Rotavirus Infections / diagnosis*
  • Sensitivity and Specificity
  • Transcription, Genetic

Substances

  • DNA Primers
  • DNA, Viral
  • RNA, Viral