High throughput PCR detection of HCV based on semiautomated multisample RNA capture

J Virol Methods. 1997 Mar;64(2):147-59. doi: 10.1016/s0166-0934(96)02153-2.

Abstract

An amplification procedure based on a semiautomated 60-sample RNA capture, including combined reverse transcription/polymerase chain reaction (RT-PCR) and nested PCR/Taqman amplicon detection. is described. It can be completed within a working day and is suitable for the development of a fully automated system. HCV RNA-specific capture is independent of the sequence variations as it targets the poly(U) tract commonly present at the 3'-end of the HCV genome (U-capture). The type specificity of the assay determined in a panel of 56 confirmed HCV antibody-positive samples (genotypes 1-6) was slightly better when compared to a commercial assay. The sensitivity evaluated on serial dilutions of representative samples was equal for genotypes 1, 2, 5, 6, or increased for genotypes 3 and 4 with the U-capture assay.

MeSH terms

  • Automation
  • Hepacivirus / genetics
  • Hepacivirus / isolation & purification*
  • Hepatitis C / blood
  • Hepatitis C / virology
  • Humans
  • Poly U
  • Polymerase Chain Reaction / methods*
  • Prospective Studies
  • RNA, Viral / analysis
  • Sensitivity and Specificity

Substances

  • RNA, Viral
  • Poly U