Objective To evaluate the performance of a DNA microarray method for detecting HBV antiviral drug-resistant mutations. Methods Two hundred and twenty four serum samples from patients with CHB were tested in parallel by DNA microarray and direct sequencing for the mutations within the HBV reverse transcriptase (rt) region, which included rtL180, rtA181, rtM204 and rtN236. Samples with discrepant results were retested by clonal sequencing. Results Complete concordance between DNA microarray and direct sequencing results was observed in 214 out of 224 samples (95. 5% ). The presence of mixed viral populations in the other 10 samples detected by DNA microarray but not by direct sequencing was confirmed by clonal analysis. The DNA microarray could detect minor viral populations which constituted 5.0%-15. 0% of the total viral load. Conclusion DNA microarray is highly consistent with direct sequencing in detecting HBV mutations conferring drug resistance and more sensitive in detecting mixed mutant and wild-type sequences than direct sequencing, which makes it a useful tool for early detection of drug resistance early.%目的 评价一种新研制的DNA微阵列芯片HBV基因型耐药检测试剂盒的临床应用性能.方法 收集2008年12月至2010年6月224份CHB患者血清标本,提取HBV DNA,应用DNA微阵列芯片法和直接测序法平行检测HBV逆转录酶区(rt区)位点rtL180、rtA181、rtM204和rtN236的耐药突变.对结果不一致的标本进行克隆测序,以验证检测的准确性.结果 芯片法和直接测序法均成功检测224份标本全部896个位点的耐药基因型,214份标本结果完全一致,其中82份检测到突变,132份为野生型HBV.2种方法检测HBV耐药突变结果的完全符合率为95.5%(214/224).其余10份标本只用芯片法检测到突变:2份rtL180M突变、2份rtA181V突变、3份rtM204I突变、2份rtM204V突变、1份rtN236T突变,但是直接测序未检测到相应突变.包含突变序列的克隆所占比例为5.0%～15.0%.经克隆测序验证结果与芯片法完全一致.结论 DNA微阵列芯片法检测HBV耐药突变与直接测序法相比符合率高,并可检出低比例耐药突变毒株,有助于早期提示耐药的发生.
展开▼
