Kamisango K, Kamogawa C, Sumi M, Goto S, Hirao A, Gonzales F, Yasuda K, Iino S
Diagnostics Research Laboratories, Chugai Diagnostics Science Co., Ltd., 3-41-8 Takada, Toshima-ku, Tokyo 171, Japan.
J Clin Microbiol. 1999 Feb;37(2):310-4. doi: 10.1128/JCM.37.2.310-314.1999.
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.
我们开发了一种灵敏的定量检测方法，该方法采用转录介导扩增和杂交保护分析法来检测血清中的乙型肝炎病毒（HBV）DNA。转录介导扩增在单管中进行。杂交保护分析在微量滴定板中使用具有不同比活性的两种探针进行，以获得较宽的检测范围。结果，该检测方法的检测范围为5×10³至5×10⁸基因组当量（GE）/毫升，并且在对数尺度上具有良好的定量准确性。一次中等规模的手工检测可在5小时内完成。通过该检测方法对临床样本中HBV DNA量的测量显示，在各种疾病状况下其含量分布广泛（超过5个对数，从约5×10³至5×10⁸ GE/毫升）。还表明，一名慢性肝炎患者的HBV DNA量变化很大，在长期监测期间范围超过5个对数。我们的检测方法有潜力用于监测和确定HBV患者及携带者的预后，尤其是在干扰素治疗期间。
