A rapid and quantitative solution hybridization method for detection of HBV DNA in serum.
A sensitive and convenient solution hybridization technique was adapted for the semiquantitative detection of hepatitis B virus DNA in serum. The assay utilizes 35S-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 microliters 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.
Jalava
T
T
Orion Corporation, Orion Pharmaceutica, Biotechnology, Helsinki, Finland.
Ranki
M
M
Bengtström
M
M
Pohjanpelto
P
P
Kallio
A
A
D004279:DNA, Viral / Q000097:blood*; D006509:Hepatitis B / Q000382:microbiology; D006514:Hepatitis B Surface Antigens / Q000097:blood; D006515:Hepatitis B virus / Q000302:isolation & purification*; D006801:Humans; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D012680:Sensitivity and Specificity
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens