Comparative analysis of different PCR techniques for detection of HCV in hepatocellular carcinoma patients

ABSTRACT

Three different isolation procedures and a primer set from the 5-TUR were compared using 3 variable PCR techniques [direct PCR, semi-nested PCR and single tube RT-PCR]. This was applied for detection of HCV- RNA in sera from 30 hepatocellular carcinoma patients positive for HCV antibodies by both EIA and immunoblotting techniques. It was found that RNA extraction with silica [Boom et al., 1990] and semi-nested PCR technique were the most sensitive procedures. By this combined method the study was able to detect 20 copies of the in vitro transcribed RNA compared to 50 copies and 100 copies using 1 tube RTPCR and direct PCR techniques, respectively