Lai V C, Guan R, Wood M L, Lo S K, Yuen M F, Lai C L
Department of Medicine, University of Hong Kong, Hong Kong.
J Clin Microbiol. 1999 Jan;37(1):161-4. doi: 10.1128/JCM.37.1.161-164.1999.
A nucleic acid photo-cross-linking technology was used to develop a direct assay for the quantification of hepatitis B virus (HBV) DNA levels in serum. Cross-linker-modified DNA probes complementary to the viral genomes of the major HBV subtypes were synthesized and used in an assay that could be completed in less than 6 h. The quantification range of the assay, as determined by testing serial dilutions of Eurohep HBV reference standards and cloned HBV DNA, was 5 x 10(5) to 3 x 10(9) molecules of HBV DNA/ml of serum. Within-run and between-run coefficients of variation (CVs) for the assay were 4. 3 and 4.0%, respectively. The assay was used to determine HBV DNA levels in 302 serum samples, and the results were compared to those obtained after testing the same samples with the Chiron branched-DNA (bDNA) assay for HBV DNA. Of the samples tested, 218 were positive for HBV DNA by both assays and 72 gave results below the cutoff for both assays. Of the remaining 12 samples, 10 were positive for HBV DNA by the cross-linking assay only; the 2 other samples were positive by the bDNA assay only. Twenty-eight samples had to be retested by the bDNA assay (CV, >20% between the results obtained from the testing of each sample in duplicate), whereas only three samples required retesting by the cross-linking assay. The correlation between the HBV DNA levels, as measured by the two tests, was very high (r = 0.902; P = 0.01). We conclude that the cross-linking assay is a sensitive and reproducible method for the detection and quantification of HBV DNA levels in serum.
一种核酸光交联技术被用于开发一种直接检测血清中乙型肝炎病毒（HBV）DNA水平的方法。合成了与主要HBV亚型病毒基因组互补的交联剂修饰DNA探针，并用于一种可在不到6小时内完成的检测方法。通过检测欧洲肝炎病毒（Eurohep）HBV参考标准品和克隆的HBV DNA的系列稀释液确定，该检测方法的定量范围为每毫升血清中5×10⁵至3×10⁹个HBV DNA分子。该检测方法的批内和批间变异系数（CV）分别为4.3%和4.0%。该检测方法用于测定302份血清样本中的HBV DNA水平，并将结果与用Chiron分支DNA（bDNA）检测法检测相同样本后获得的结果进行比较。在检测的样本中，两种检测方法均显示218份样本的HBV DNA呈阳性，72份样本的检测结果均低于两种检测方法的临界值。在其余12份样本中，10份仅通过交联检测法显示HBV DNA呈阳性；另外2份样本仅通过bDNA检测法呈阳性。28份样本必须通过bDNA检测法重新检测（CV，对每个样本进行重复检测所获得的结果之间>20%），而只有3份样本需要通过交联检测法重新检测。两种检测方法所测得的HBV DNA水平之间的相关性非常高（r = 0.902；P = 0.01）。我们得出结论，交联检测法是一种用于检测和定量血清中HBV DNA水平的灵敏且可重复的方法。
