核苷(酸)类药治疗慢性乙肝患者HBV逆转录酶区基因耐药的变异分析

Analysis of drug-resistant mutation in the RT region of hepatitis B virus P gene in patients treated with nucleoside/nucleotide analogs

目的:了解核苷(酸)类药物治疗慢性乙型肝炎过程中HBV耐药变异特点。方法:收集163份长期核苷(酸)类药物治疗的慢性乙型肝炎患者血清标本,用半巢式聚合酶链反应-直接测序法获得HBV全长逆转录酶区序列,用生物信息学技术筛查该区内6个经典耐药变异位点并鉴定基因型,分析慢性乙型肝炎核苷(酸)类药物耐药特征及与HBV基因型关系。结果:163份慢性乙型肝炎患者血清标本中有64份(占39.26%)发现了已知耐药位点变异。拉米夫定(1amivudine,LAM)治疗组耐药性变异比例高于阿德福韦(adefovir dipivoxil,ADV)治疗组。单用LAM组和单用ADV组发现已知耐药位点变异比例分别为39.40%和12.90%。19例LAM和ADV序贯治疗的患者中出现5例(占26.32%)多药耐药变异特征,而LAM与ADV联合治疗的16例患者中未发现后续ADV耐药变异。耐药变异与HBV基因型相关性分析显示,存在RT基因耐药位点变异的单用LAM组、单用ADV组、LAM和ADV序贯治疗组和LAM治疗后加用ADV组间的HBV基因型比例差异无统计学意义(P>0.05)。结论:慢性乙型肝炎长期核苷(酸)类药物治疗,耐药变异复杂,对LAM耐药的患者,改用ADV比加用ADV更易导致多重耐药变异。核苷(酸)类药物耐药变异与HBV基因型别无关。

Objective： To analyze the feature of drug-resistant mutation in the RT region of hepatitis B virus（HBV） P gene in patients treated with nucleoside or nucleotide analogs.Methods： Full-length reverse transcriptase region of HBV DNA was amplified by semi-nested polymerase chain reaction from 163 chronic hepatitis B（CHB） patient treated with nucleoside or nucleotide analogs.The PCR products were directly sequenced.Then,six classical anti-viral drug-resistant mutation sites were screened by bioinformatics.The relationship between the drug-resistant mutation and HBV genotypes was further analyzed.Results： Known drug-resistant mutations were found in 64 cases（39.26%） among 163 samples.The incidence of drug resistance in patients treated with lamivudine was higher than those treated with adefovir.The proportions of known drug-resistant mutations in the group of single-dose lamivudine and adefovir treatment were 39.40% and 12.90%,respectively.The incidence of adefovir-resistant mutation in patients sequentially treated lamivudine and adefovir was 26.32%（5/19）,whereas no patients treated with lamivudine plus adefovir in combination shown the feature of adefovir-resistant mutation.The distribution of HBV genotype in the groups of single-dose lamivudine,single-dose adefovir,sequential or combination treatment of lamivudine and adefovir shown no significantly different.Conclusion： During the treatment of nucleotide analogue in CHB patients,the drug-resistant mutation was complex.In lamivudine resistant patients,switching to adefovir was more incidence of multi-drug resistance than combination of lamivudine and adefovir.The relationship of drug-resistant mutations and genotypes of HBV showed no significance.