High throughput PCR detection of HCV based on semiautomated multisample RNA capture.
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.
Petrik
J
J
Department of Haematology, University of Cambridge, UK.
Pearson
G J
GJ
Allain
J P
JP
D001331:Automation; D016174:Hepacivirus / Q000235:genetics / Q000302:isolation & purification*; D006526:Hepatitis C / Q000097:blood / Q000821:virology; D006801:Humans; D011072:Poly U; D016133:Polymerase Chain Reaction / Q000379:methods*; D011446:Prospective Studies; D012367:RNA, Viral / Q000032:analysis; D012680:Sensitivity and Specificity
D012367:RNA, Viral; D011072:Poly U