Quantitative detection of hepatitis B virus by transcription-mediated amplification and hybridization protection assay.
We have developed a sensitive and quantitative assay using transcription-mediated amplification and hybridization protection assay for the detection of hepatitis B virus (HBV) DNA in serum. The transcription-mediated amplification was carried out in a single tube. The hybridization protection assay was carried out in a microtiter plate with two probes with different specific activities to obtain a broad detection range. As a result, the assay had a detection range of 5 x 10(3) to 5 x 10(8) genome equivalents (GE)/ml and good quantitative accuracy on a logarithmic scale. A moderately sized manual assay run can be completed within 5 h. Measurements of the amounts of HBV DNA in clinical samples by the assay showed the amounts under various disease conditions to be widely distributed (more than 5 logs, from approximately 5 x 10(3) to 5 x 10(8) GE/ml). It was also shown that the amount of HBV DNA in one chronic hepatitis patient varied widely, with a range of more than 5 logs during long-term monitoring. Our assay has the potential to be used to monitor and determine the prognosis of HBV patients and carriers, especially during interferon treatment.
Kamisango
K
K
Diagnostics Research Laboratories, Chugai Diagnostics Science Co., Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171, Japan. kamisangokii@chugai-pharm.co.jp
Kamogawa
C
C
Sumi
M
M
Goto
S
S
Hirao
A
A
Gonzales
F
F
Yasuda
K
K
Iino
S
S
D006528:Carcinoma, Hepatocellular / Q000821:virology; D002353:Carrier State / Q000821:virology; D004279:DNA, Viral / Q000097:blood*; D005784:Gene Amplification; D006509:Hepatitis B / Q000821:virology*; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification*; D019694:Hepatitis B, Chronic / Q000821:virology; D006801:Humans; D008103:Liver Cirrhosis / Q000821:virology; D008113:Liver Neoplasms / Q000821:virology; D009693:Nucleic Acid Hybridization; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014158:Transcription, Genetic; D014766:Viremia / Q000821:virology
D004279:DNA, Viral; D011933:Reagent Kits, Diagnostic