乙型肝炎病毒阿德福韦酯耐药株基因组分析

Cloning and analysis of complete sequence of Adefovir dipivoxil resistant hepatitis B virus

目的:分析乙型肝炎病毒(hepatitis B virus,HBV)阿德福韦酯耐药株的基因序列。方法:从已发生阿德福韦酯临床耐药的慢性乙型肝炎患者血清中提取HBV DNA,设计保守引物,采用高保真聚合酶对耐药株HBV DNA进行全序列扩增,通过克隆技术构建PGEM-HBV重组质粒,测序后进行序列分析。结果:获得阿德福韦酯耐药的HBV DNA全长序列克隆,序列长度为3 261 bp,序列分析发现在逆转录区有rt A181V位点突变。结论:阿德福韦酯耐药株HBV DNA全长序列克隆的成功构建,为阿德福韦酯临床耐药性研究提供参考。

Objective： To analyze genomic sequences the full-length DNA of Adefovir-resistant hepatitis B virus（HBV）strains. Methods： HBV DNA in serum was extracted from an Adefovir dipivoxil resistant patient with chronic hepatitis B by the conservative primers,PCR was performed for complete genome amplification with high-fidelity enzyme,and the products were cloned to the plasmid for sequencing and analysis. Results： The HBV DNA full-length sequence with HBV Adefovir resistance was obtained,which composed of 3 261 bp. A mutation of rt A181 V was found in the reverse transcription through sequence analysis. Conclusion： A complete sequence of HBV DNA from an Adefovir dipivoxil resistant was obtained successfully,which will be helpful in the development of new drug for Adefovir dipivoxil resistance.