慢性HBV感染患者核苷类药物治疗后相应突变位点检测分析

Detection and analysis of nucleoside drug-resistant mutation in patients with chronic hepatitis B

目的:通过基因测序检测分析拉米夫定(LAM)、阿德福韦(ADV)、恩替卡韦(ETV)、替比夫定(LdT)四种核苷类似物耐药者HBV多聚酶区基因主要耐药突变的分布情况。方法:采用实时荧光定量PCR方法对253例LAM,78例ADV和9例ETV以及6例LdT耐药患者血清HBV多聚酶区进行扩增,对PCR产物进行直接测序。结果:253例LAM耐药患者中有16种突变类型,其中以rtM204I突变最多见,为101例(39.92%)。78例ADV耐药患者中有7种突变类型,其中以rtA181V/T/S突变最多见,为47例(60.25%)。9例ETV耐药患者中有3种突变类型,其中以rtT184A/G/I/S突变最多见,为6例(66.66%)。6例LdT耐药患者中有2种突变类型。结论:四种核苷类似物耐药株的氨基酸突变复杂多样,建议抗病毒治疗失败后参考基因型耐药检测结果更换药物。

Objective：To analyze the drug resistance mutation distribution of HBV polymerase region gene of four nucleoside analogues including lamivudine（LAM）,adefovir dipivoxil（ADV）,entecavir（ETV） and telbivudine（LdT）,using PCR product direct sequencing method.Methods： The HBV polymerase region of serum samples of chronic hepatitis B（CHB） patients were amplified with Real-time PCR,including 253 cases of LAM-resistance,78 cases of ADV-resistance,9 cases of ETV-resistance and 6 cases of LdT-resistance,and the PCR products were directly sequenced.Results： Sixteen kinds of HBV polymerase region gene mutations were detected within 253 LAM-resistance CHB patients,and rtM204I was the most common mutation（n=101,39.92%）.Seven kinds of mutations were detected within 78 ADV-resistance CHB patients and the major mutation was rtA181V/T/S（n=47,60.25%）.Three kinds of mutations were detected in 9 cases of ETV-resistance patients and the most common mutation was rtT184A/G/I/S（n=6,66.66%）.Two kinds of mutations were observed in 6 cases of LdT-resistance patients.Conclusion： Given diverse amino mutations related with four nucleoside analogues resistance,the drug resistant genotype testing in patients with failure antiviral treatment maybe a valuable reference for drug selection.