HEPATITIS-B VIRUS;
SOLUTION HYBRIDIZATION;
QUANTIFICATION OF DNA;
DIAGNOSTICS;
D O I:
10.1016/0166-0934(92)90148-7
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
A sensitive and convenient solution hybridization technique was adapted for the semiquantitative detection of hepatitis B virus DNA in serum. The assay utilizes S-35-isotope as label and biotin-avidin interaction for collection of the hybrids onto microtitre plate wells. Results are obtained as numerical values, which allow quantification. 10(6) molecules of HBV DNA/ml serum could be detected by a 3-h hybridization followed by a 2-h collection reaction. By analyzing 500-mu-l of serum, 30 (88%) of 34 patient sera positive for HBeAg were also positive for HBV DNA and 17 HBsAg-positive sera, 16 of which were anti-HBe-positive, were DNA-negative. The amount of HBV DNA varied from 5 x 10(6) to 3 x 10(9) molecules/ml. The solution hybridization method which was developed allows fast and accurate quantification of HBV DNA in serum providing an estimate of the virus titre.