Quantitative detection of hepatitis B virus by transcription-mediated amplification and hybridization protection assay

We have developed a sensitive and quantitative assay using transcription-me diated amplification and hybridization protection assay for the detection o f hepatitis B virus (HBV) DNA in serum. The transcription-mediated amplific ation was carried out in a single tube. The hybridization protection assay was carried out in a microtiter plate with two probes with different specif ic 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 manu al assay run can be completed within 5 h, Measurements of the amounts of HB V DNA in clinical samples by the assay showed the amounts under various dis ease conditions to be widely distributed (more than 5 logs, from approximat ely 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 t han 5 logs during long-term monitoring. Our assay has the potential to be u sed to monitor and determine the prognosis of HBV patients and carriers, es pecially during interferon treatment.