乙肝病毒基因分型检测芯片的构建及初步应用

Construction and clinical application of hepatitis B virus genotyping microarray

目的:构建乙肝病毒(HBV)常见基因型的分型芯片,并应用分型芯片调查HBV基因型分布。方法:应用生物信息学软件针对乙肝病毒4种常见基因型的特征区域和保守区域分别设计分型探针和通用探针,在优化条件下通过重复杂交及分型测序检验芯片的重复性与可靠性后应用于临床,对150例不同类型的乙肝患者进行HBV基因分型。结果:分型探针和通用探针检测重现率分别为94.7%及100%,反馈的测序结果与芯片分型结果完全一致;经芯片分型发现辽宁省HBV基因型分别为C型占68.7%,B型占22.7%,B、C混合型占8.7%,未发现A、D型。结论:基因芯片技术为乙肝病毒基因分型提供了快速准确的检测方法,辽宁地区HBV基因型分布以B、C两型为主,C型占主要优势。

Objective：To construct a DNA microarray for quick detection and identification of hepatitis B virus （HBV） genotypes,and to study the distribution of HBV genotypes of patients with hepatitis in Liaoning Province by using this microarray. Methods：The genotyping probes and universal probes were designed based on the specific and conserved regions of HBV genome,respectively. The reproducibility and reliability of the microarray were tested by repeated hybridization and sequencing,and the genotypes of HBV DNA extracted from serum samples of 150 patients with hepatitis B were detected by using this microarray. Results：The reproducibility rates of genotyping probe and universal probe were 94.6% and 100%,respectively,and the sequence analysis was consistent with the result of microarray assay. The percentages of genotypes C,B,and BC were 68.6%,22.6%,and 8.6%,respectively. Conclusion：This DNA microarray can be used for quick HBV genotyping. The HBV genotypes in the patients with hepatitis in Liaoning Province are mainly genotypes B and C,and genotype C is predominant.