Detection of hepatitis C virus genome in human serum by multi‐targeted polymerase chain reaction

M. Okamoto, M. Baba, E. Kodama, K. Sekine, T. Takagi, R. Kasukawa, S. Shigeta

Research output: Contribution to journalArticlepeer-review

8 Citations (Scopus)

Abstract

A multi‐targeted “hemi‐nested” PCR (M‐PCR) assay in which the primer pairs derived from the 5′ non‐coding (5′NC) and the nonstructural protein 3 (NS3) regions of HCV genome were concurrently used for amplification in order to compare the sensitivity and specificity of polymerase chain reaction (PCR) with different primer pairs in detecting hepatitis C virus (HCV) genome. Sera from patients with virus‐associated liver diseases were examined for the presence of HCV RNA by the M‐PCR method following reverse transcription to cDNA. The amplified products derived from both the 5′NC and the NS3 regions were detected in 28 (70%) of the 40 HCV RNA‐positive samples. However, 12 samples (30%) were devoid of the signal of NS3‐derived product. Sensitivity tests using serial dilutions of HCV RNA revealed that the 5′NC‐derived band was still detectable in the 105‐fold diluted sample by the M‐PCR method, yet the NS3‐derived band could hardly be detected in the 104‐fold diluted sample. Thus, as previously demonstrated by a single‐targeted “nested” PCR assay, the present study using the M‐PCR assay has clearly shown that the 5′NC‐derived primers are more sensitive and specific than the NS3‐derived primers in detecting HCV RNA.

Original languageEnglish
Pages (from-to)6-10
Number of pages5
JournalJournal of Medical Virology
Volume41
Issue number1
DOIs
Publication statusPublished - 1993 Sep
Externally publishedYes

Keywords

  • HCV
  • PCR
  • hepatitis C virus
  • human serum
  • multiple targets
  • polymerase chain reaction

ASJC Scopus subject areas

  • Virology
  • Infectious Diseases

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