Quantitative Measurement of PCR-amplified Hepatitis B Virus DNA Using Enzyme Immunoassay

A quantitative polymerase chain reaction enzyme immunoassay (PCR-EIA) for the detection of hepatitis B virus (HBV) DNA in serum samples was established and evaluated by testing 120 samples in compared with detection of PCR products by agarose gel electrophorsis with ethidium bromide staining (PCR-EB). Of these, 43 samples (43 cases) were collected from hepatitis individual carriers with hepatitis B surface antigen-positive and hepatitis B e antigen-positive, 19 samples (19 cases) were collected from pretreatment hepatoma patients and 58 samples (18 cases) were collected from follow-up treatment hepatoma patients. A sensitivity of PCR-EIA was 8.64 x 10 2 copies/ml whereas that of PCR-EB was 3.48 x 10 6 copies/ml in standard HBV DNA. The detection level of HBV DNA for PCR-EIA was in the range of 8.64 x 10 2 to 1.74 x 10 6 copies/mI. For hepatitis carriers, detection of HBV DNA was 97.7% by PCR-EIA and 95.3% by PCR-EB. 3 of 58 samples from follow-up cases were negative for HBV DNA by PCR-EB but all of them were positive by PCR-EIA. The results indicated that the PCR-EIA is more sensitive than the PCR-EB and it can be applied for diagnosis of HBV infection and prognosis of disease progression. Key words: HBV DNA, PCR-EIA, PCR-EB