Direct polymerase chain reaction for detection of hepatitis B, C and G virus genomes from serum without nucleic acid extraction - simple, rapid and highly sensitive method

Although hepatitis C virus (HCV) detection by polymerase chain reaction (PC R) assay is now the standard, extensive clinical application has been thwar ted by the troublesome procedure, long reaction lime and potential for cont amination. To overcome these problems, we carried out a PCR assay for the d etection of HCV, hepatitis G virus (HGV) and hepatitis B virus (HBV) genome s directly from serum samples without any nucleic acid extraction (direct P CR). The sensitivity of this assay was one chimpanzee-infectious dose of HC V and a 10(-1) chimpanzee-infectious dose of HBV. This result was similar t o the sensitivity determined by the conventional PCR. Furthermore, when the detection rate of these genomes in serum samples from chimpanzees and huma ns is compared, the results matched completely between two different PCR as says. The whole process, including the reverse transcriptase reaction and s econd round PCR, can be completed within 6 h by the combination of the dire ct PCR and one-step PCR assay. Our findings indicate that this method is si mple, rapid and highly sensitive and could be useful for the screening of b lood-borne hepatitis virus infections using serum samples. (C) 1998 Elsevie r Science Ireland Ltd. All rights reserved.