Detection of negative-stranded hepatitis C virus RNA using a novel strand-specific reverse transcription-polymerase chain reaction

Tetsuya Mizutani, Masanori Ikeda, Satoru Saito, Kazuo Sugiyama, Kunitada Shimotohno, Nobuyuki Kato

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

We developed a novel single-tube reverse transcription-polymerase chain reaction (RT-PCR) for the specific detection of negative-stranded hepatitis C virus (HCV) RNA. By using in vitro synthesized positive- and negative-stranded HCV RNAs, it was demonstrated that as few as 50 copies of negative-stranded HCV RNA could be specifically detected with a set of primers that amplify a 232-base pair sequence unique to the 5'-non-coding region of HCV RNA, while 108 copies of positive-stranded HCV RNA were not detected. In addition, we demonstrated that this method allows the detection as few as 100 copies of negative-stranded HCV RNA even with the coexistence of a 100-fold excess of positive-stranded HCV RNA. Furthermore, with this method, negative-stranded HCV RNA was detected in RNAs from liver biopsy specimens obtained from patients with chronic hepatitis C, but not in RNAs from HCV-positive sera. Copyright (C) 1998 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)209-214
Number of pages6
JournalVirus Research
Volume53
Issue number2
DOIs
Publication statusPublished - Feb 1 1998
Externally publishedYes

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Keywords

  • 5'-non-coding region
  • HCV replication
  • Hepatitis C virus
  • Negative strand
  • Polymerase chain reaction

ASJC Scopus subject areas

  • Cancer Research
  • Molecular Biology
  • Virology

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