To estimate the clinical efficiency of digoxigenin(DIG) labeled DNA probe for the detection of hepatitis B virus(HBV) DNA, 149 HBs Ag positive and 15 normal serum samples were tested by DNA dot blot hybridization using 32P-dCTP-labeled HBV DNA
probe and
DIG-dUTP-labeled HBV DNA probe. Serological markers were tested by ELISA.
The sensitivity of 32P-dCTP-labeled HBV DNA probe in detecting HBV DNA was 0.5㎍, that of DIG-dUTP-labeled HBV DNA probe labeled by random primed labeling method was 4㎍, and the sensitivity of DIG-dUTP-labeled HBV DNA probe labeled by polymerase
chain
reaction(PCR) was 0.5㎍.
Expire time of 32P-dCTP-labeled HBV DNA probe was within a within a week, that of DIG-dUTP-labeled HBV DNA probe labeled by random primed labeling method was 4 to 6 months, and that of DIG-dUTP-labeled HBV DNA probe labeled by PCR was 2 to 4
weeks.
Time needed for autoradiography was 24-48 hours, but in the case of DIG-labeled probe, visualization of the result needed only 6 hours.
The relationship between serum HBV DNA positivity by DIG-labeled probe and serological markers positivity was evaluated, Twenty-seven(79.4%) of 34 patients positive for both HBs Ag and Hbe Agf were positive for HBV DNA compared with 25(21.7%) of
115 HBS
Ag positive but Hbe Ag negative individuals. There as a statistically significant correlation between the positivity of HBV DNA in serum and the presence or absence of Hbe Ag. None of the 15 HBs Ag and Hbe Ag negative individuals tested was
positive for
HBV DNA. But the detection rate of HBV DNA in the cases of Hbe Ag negative was as high as 21.7%(25/115), so we concluded that dot blot hybridization technie using DIG-labeled HBV DNA probe could be very efficiently applied for both diagnosing HBV
infection and evaluating the infectivity of the HBs Ag positive sera.
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