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

Fingerprint

Viruses
Hepacivirus
RNA
Polymerase Chain Reaction
Renilla Luciferases
Nucleotides
Lactoferrin
Virus Diseases
Virus Replication
Antiviral Agents
Hepatocytes
Sensitivity and Specificity

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.

A reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus RNA. / Nozaki, Akito; Naganuma, Atsushi; Nakamura, Takashi; Tanaka, Katsuaki; Sekihara, Hisahiko; Kato, Nobuyuki.

In: Acta Medica Okayama, Vol. 54, No. 6, 2000, p. 253-257.

Research output: Contribution to journalArticle

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, vol. 54, no. 6, pp. 253-257.
Nozaki, Akito ; Naganuma, Atsushi ; Nakamura, Takashi ; Tanaka, Katsuaki ; Sekihara, Hisahiko ; Kato, Nobuyuki. / A reliable internally controlled RT-nested PCR method for the detection of hepatitis C virus RNA. In: Acta Medica Okayama. 2000 ; Vol. 54, No. 6. pp. 253-257.
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