A ribonuclease protection assay for the direct detection and quantitation of hepatitis C virus RNA

Nafees Ahmad, I. Ken Kuramoto, Bahige M. Baroudy

Research output: Contribution to journalArticle

7 Scopus citations

Abstract

A ribonuclease protection assay (RPA) was developed for the direct detection and quantitation of HCV RNA in infected patients' sera or plasma using HCV [32P]RNA from the conserved 5′-untranslated region (5′-UTR) as a probe. We were able to directly detect the presence of HCV RNA by RPA in several infected patients' samples. The viremic status of HCV infected patients with indeterminate recombinant immunoblot assay (RIBA II) was also determined by this assay. Polymerase chain reaction (PCR) was also performed on all these samples and were found to be positive with a concordance of 100% between the results of PCR and RPA. The RPA was able to detect approximately 1 pg of HCV RNA. A limited sequence heterogeneity among HCV isolates was also observed by this assay, suggesting that this may be a faster method of detecting heterogeneous HCV sequences in patients' samples. This simple and specific method could be used to quantitate HCV RNA in order to better determine viremia and follow the course of HCV infection especially when RIBA II results are indeterminate.

Original languageEnglish (US)
Pages (from-to)233-244
Number of pages12
JournalClinical and Diagnostic Virology
Volume1
Issue number4
DOIs
StatePublished - Oct 1993
Externally publishedYes

Keywords

  • 5′-Untranslated region
  • HCV
  • Polymerase chain reaction
  • Ribonuclease protection assay
  • Viremia

ASJC Scopus subject areas

  • Virology

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