核酸序列测定技术分析HBV耐药基因与亚型鉴定

Detection of Drug Resistance Gene and Subgenotype of HBV with Nucleic Acid Sequencing Method

目的:用荧光标记核酸序列测定技术分析患者携带的HBV基因型,判断所携带的HBV亚型以及治疗过程中是否产生耐药性。方法:采用PCR产物直接进行核酸序列分析,使用计算机模拟核酸杂交技术分析HBV基因的亚型与耐药基因。结果:扩增产物的核酸序列分析表明,设计的引物序列在确定的条件下可以特异性的扩增HBV相应的基因片段。患者耐药基因分析结果分别为HBVYVDD变异和YIDD变异;病毒亚型分析结果提示患者携带的HBV均为adr亚型。结论:本方法能够准确判定HBV的YVDD与YIDD变异及其它类型突变,并确定患者携带HBV的基因型,能够准确、客观地反映出病原体的耐药性,可以作为合理用药的重要参考依据。

Objective To identify sub - genotype and presence of drug resistance mutation during lamivudine therapy with fluorescence labeled nucleic acid sequencing method. Methods In six lamivudine resistant patients, PCR products were used for nucleic acid sequencing, computer simulation of nucleic acid hybridization were used to assay HBV sub - genotype and drug resistance gene. Results Specific HBV gene fragment was obtained with self - designed primer. Drug resistance genes in patients were of YVDD variants and YIDD variants. The HBV sub - genotype was adr genotype in all patients. Conclusion Nucleic acid sequencing can accurately identify YVDD variants, YIDD variants and other mutants, and detect HBV genotype, which is important for selecting proper drug for treatment.