Quantitative method of intracellular hepatitis C virus RNA using LightCycler PCR

Akito Nozaki, Nobuyuki Kato

Research output: Contribution to journalArticlepeer-review

13 Citations (Scopus)

Abstract

Based on recent LightCycler techniques developed for the quantitation of serum HCV RNA, we have developed a quantitative method for the intracellular hepatitis C virus (HCV) RNA using LightCycler PCR. A simple real-time PCR assay, based on the SYBR Green I dye and LightCycler fluorimeter and with no probe requirement, is described. In the presence of 0.5 μg of cellular RNA, it was demonstrated that as few as 25 copies of HCV RNA could be specifically detected with a set of primers that amplify a 144-base pair sequence unique to the 5'-noncoding region of HCV RNA. We demonstrated that this method was useful for the evaluation of antiviral reagents using HCV-infected human cultured cells.

Original languageEnglish
Pages (from-to)107-110
Number of pages4
JournalActa medica Okayama
Volume56
Issue number2
Publication statusPublished - 2002

Keywords

  • Hepatitis C virus
  • LightCycler
  • Real-time PCR

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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