DETECTION OF HEPATITIS-C VIRUS BY UNBALANCED POLYMERASE-CHAIN REACTION, HYBRIDIZATION TO SYNTHETIC OLIGONUCLEOTIDES AND CAPILLARY ELECTROPHORESIS

Polymerase chain reaction (PCR) has been reported as one of the most e fficient techniques to detect hepatitis C virus (HCV) RNA. The determi nation of the specificity of PCR products is usually based on 'nested' PCR, Southern blotting and hybridization of the amplified DNA to radi oactive oligonucleotide probes recognizing sequences comprised between the PCR primers. The recent introduction of capillary electrophoresis (CE) to analyse DNA fragments and PCR products appears to be very int eresting because this technology is rapid, reproducible, sensitive and could be suitable to detect DNA/DNA and DNA/RNA hybrids. We demonstra te that specific hybridization of an HCV oligonucleotide probe to sing le stranded HCV-DNA obtained by unbalanced PCR is detectable by capill ary electrophoresis, therefore enabling a one-step, non-radioactive pr otocol to demonstrate the specificity of amplification of HCV sequence s by PCR.