Supersensitive detection method of HBV DNA using polymerase chain reaction

Abstract

The principle of polymerase chain reaction (PCR) is to amplify the DNA exponentially by repeating the polymerase reaction. We detected the HBV DNA in serum specimens applying PCR method. For determination of the sensitivity of this method, serially diluted HBV DNA (1∼10-7pg) were examined. Cloned HBV DNA equivalent to one virus genome (10-6pg) was detectable by ethidium bromide staining after 50 cycles of PCR. The amplified DNA with PCR was proven to have the homology with HBV DNA by Southern blot. In serum specimens, HBV DNA was detected in 5 of 5 HBeAg seropositive, in 2 of 2 HBe Ag/Ab seronegative, in 8 of 10HBeAb seropositive, but in none of 6 HBsAg seronegative cases. Small amount of HB virus seems to present in sera of many HBeAg seropositive patients. © 1989, The Japan Society of Hepatology. All rights reserved.