Optimization, assessment, and proposed use of a direct nested reverse transcription-polymerase chain reaction protocol for the detection of hepatitis C virus

Mohamed Abdel-Hamid, Daniel C. Edelman, W. Edward Highsmith, Niel T. Constantine

Research output: Contribution to journalArticle

68 Scopus citations

Abstract

Objectives: To compare the performance of reverse transcription followed by the polymerase chain reaction (RT-PCR), without RNA purification, with the performance of classic protocols. Study Design/Methods: Direct and classic techniques were used to test three groups of samples: six hepatitis C virus (HCV) seroconversion panels (n = 90), a HCV RNA reference panel (n = 26), and serial dilutions of four HCV-positive sera (n = 24). These methods were then applied sequentially through a clinical diagnostic algorithm to test 268 samples from high-risk patients. Results: For the three groups of samples, we found a 94% concordance between direct and purified RT-PCR methods. For the detection of HCV RNA in clinical samples, sensitivity was maximized and cost minimized using both protocols according to the proposed algorithm. Conclusions: The direct PCR method is reliable, sensitive, and can result in time and cost savings. The suggested testing algorithm can enhance sensitivity and time savings for populations with a high prevalence of infection.

Original languageEnglish (US)
Pages (from-to)58-65
Number of pages8
JournalJournal of Human Virology
Volume1
Issue number1
StatePublished - Nov 1 1997

Keywords

  • Diagnostics
  • Hepatitis
  • Hepatitis C virus
  • Molecular biology
  • Reverse transcription-polymerase chain reaction (RT-PCR) assay

ASJC Scopus subject areas

  • Virology

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