Detection of hepatitis B virus in serum using amplification of viral DNA by means of the polymerase chain reaction.
A new assay was developed for the detection of hepatitis B virus (HBV) in human serum using amplification of a short viral DNA sequence by means of the polymerase chain reaction. As little as 0.4 fg viral DNA, corresponding to about 130 genome equivalents, per ml serum could be detected after the amplification procedure. This assay detected viral DNA in a number of patients with proven or suspected chronic HBV infection who were all negative for HBV DNA in the conventional hybridisation assay. We found HBV DNA in all of six HBeAg-positive and in three of eight HBeAg-negative HBsAg carriers, as well as in all of 11 patients with chronic liver disease with antibodies against the HBV core antigen (anti-HBc) as the sole marker for HBV infection, and in three of five apparently healthy individuals showing only anti HBc. Thus, this method is an important improvement for the diagnosis of persistent HBV infections, especially in patients where a definitive serological diagnosis is not possible.
Sumazaki
R
R
Max von Pettenkofer Institute for Hygiene and Medical Microbiology, University of Munich, Federal Republic of Germany.
Motz
M
M
Wolf
H
H
Heinig
J
J
Jilg
W
W
Deinhardt
F
F
D000914:Antibodies, Viral / Q000276:immunology; D015139:Blotting, Southern; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis / Q000096:biosynthesis*; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D004587:Electrophoresis, Agar Gel; D005784:Gene Amplification; D006508:Hepatitis Antibodies / Q000032:analysis; D006509:Hepatitis B / Q000175:diagnosis; D006511:Hepatitis B Antigens / Q000276:immunology; D006515:Hepatitis B virus / Q000302:isolation & purification*; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D008722:Methods; D010957:Plasmids; D012680:Sensitivity and Specificity; D012698:Serologic Tests; D019914:Taq Polymerase
D000914:Antibodies, Viral; D015342:DNA Probes; D004279:DNA, Viral; D006508:Hepatitis Antibodies; D006511:Hepatitis B Antigens; D007106:Immune Sera; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase