SIMULTANEOUS DETECTION OF BOTH HEPATITIS-B VIRUS-DNA AND HEPATITIS-C VIRUS-RNA USING A COMBINED ONE-STEP POLYMERASE CHAIN-REACTION TECHNIQUE

Hepatitis C virus (HCV) RNA polymerase chain reaction (PCR) is widely used for diagnosis of HCV infection and evaluation of therapy, The sen sitive hepatitis B virus (HBV) DNA PCR is often reserved for detection of quantities of HBV DNA that are insufficient for hybridization. App lication of both PCR techniques is Limited by their labor-intensity, p otential for contamination, and substantial time required for analysis . To study HCV and HBV infections, occurring alone or in combination, we developed a combined one-step PCR method to detect HCV RNA and HBV DNA in a single serum specimen using oligoprimers from the HCV 5' untr anslated region and the HBV preS/S region. Specificity of the HBV and HCV PCR products was confirmed on the basis of their molecular sizes i n positive samples, Southern blot hybridization, and negative controls . The sensitivities of the combined PCR were assessed using samples co ntaining a wide range of defined amounts of HBV DNA and HCV RNA and we re comparable with those obtained with conventional HBV DNA or HCV RNA PCR methods. The sensitivity of the combined method was further valid ated by the 100% concordance between results of its HBV and HCV compon ents and those of conventional PCR methods in patients with HBV and/or HCV infections, The combined one-step HBV/HCV PCR is a sensitive, spe cific, rapid, and cost-effective method, especially suited for epidemi ological screening and clinical diagnosis of HBV and HCV infections oc curring alone or in combination.