A SIMPLE SINGLE-TUBE PROCEDURE OF PCR ASSAY FOR THE DETECTION OF HEPATITIS-C VIRUS-RNA

We developed a simple single tube procedure using the PCR (polymerase chain reaction) for the detection of hepatitis C virus RNA. The entire reaction from RNA extraction to PCR occurred in one tube; this was ma de possible by the use of more than 1,000 mu g/ml of proteinase K for RNA extraction, instead of the acid-guanidinium thiocyanate-phenol-chl oroform method. All necessary reagents were added to the tube, and PCR products were not removed from the tube until the end of PCR, althoug h opening of the tube during the procedure could not be avoided. There fore, cross-contamination which might theoretically take place during transfer of products between tubes never occurred. This method detecte d about 1 chimpanzee infectious dose of HCV in H strain plasma.