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/Tagman 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.