A reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus RNA

Akito Nozaki, Atsushi Naganuma, Takashi Nakamura, Katsuaki Tanaka, Hisahiko Sekihara, Nobuyuki Kato

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

We have developed a reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus (HCV) RNA using in vitro synthesized Renilla luciferase (Rluc) RNA as an internal control. Using this method, the 5'-noncoding region of HCV RNA (144 nucleotides) and Rluc RNA (276 nucleotides) were efficiently amplified in a single tube, and the sensitivity and specificity of this method were comparable to standard RT-nested PCR. This method was successfully performed on RNA specimens obtained from in vitro HCV-infected human hepatocyte PH5CH8 cells, which support HCV replication. In addition, we demonstrated that this method was useful for the evaluation of antiviral reagents by confirming the anti-HCV activity of bovine lactoferrin, which we previously found to be a new inhibitor of HCV infection. Therefore, this method may be useful for the studies of not only HCV but also of other viruses.

Original languageEnglish
Pages (from-to)253-257
Number of pages5
JournalActa Medica Okayama
Volume54
Issue number6
Publication statusPublished - 2000

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Keywords

  • Hepatitis C virus
  • Internal control
  • Reverse transcription-nested PCR (RT-nested PCR)

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Nozaki, A., Naganuma, A., Nakamura, T., Tanaka, K., Sekihara, H., & Kato, N. (2000). A reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus RNA. Acta Medica Okayama, 54(6), 253-257.