恩替卡韦治疗后乙型肝炎病毒多聚酶区耐药基因检测分析

Drug-resistant Genes at Hepatitis B Virus Polymerase Region during Entecavir Treatment

目的研究恩替卡韦治疗后乙型肝炎病毒(HBV)多聚酶区基因变异情况。方法收集恩替卡韦治疗后出现HBV病毒学突破患者的血清,采用聚合酶链反应(PCR)产物直接测序,分析HBV-P区基因变异发生情况。结果 29例患者中检出恩替卡韦基因型耐药19例,其中16例(84.2%,16/19)既往有拉米夫定治疗失败史;17例(58.6%,17/29)变异类型以rtL180+rtM204+rtT184为主;7例(24.1%,7/29)为B基因型,22例(75.9%,22/29)为C基因型。7例B基因型中检出恩替卡韦基因型耐药者4例(57.1%),22例C基因型中检出恩替卡韦基因型耐药者15例(68.2%),差异无统计学意义(P=0.665)。结论恩替卡韦基因型耐药多出现于拉米夫定治疗失败后序贯治疗,rtL180+rtM204+rtT184变异类型在B和C基因型的恩替卡韦耐药中较为常见。

Objective To investigate the drug-resistant genes at hepatitis B virus （HBV） polymerase region during entecavir （ETV） treatment. Methods Serum samples from chronic hepatitis B patients with viro- logic breakthrough during enticavir therapy were studied. The resistant mutation patterns in the polymerase gene of hepatitis B virus were analyzed using the polymerase chain reaction （PCR） -sequencing method. Results ETV resistance was detected from 19 out of 29 ETV-refractory patients, among whom 16 （84.2%） had a hisotry of lamivudine-refraetory. The mutation patterns were diverse, while rtL180 ＋ rtM204 ＋ rtT184 （58.6% , 17/29） was most common in patients with ETV genotype resistance. Four of 7 patients （7/29, 24. 1% ） with genotype B were detected to have ETV genotype resistance, while 15 of 22 patients （22/29, 75.9% ） with genotype C were detected to have ETV genotype resistance. The rate of ETV genotype resistance was 57. 1% （4/7） and 68.2% （15/22） in patients with genotype B and genotype C, while no statistical difference was found （P = 0. 665 ） . Conclusions ETV genotype resistance is more common in patients who have been refractory to ETV and lamivudine sequential treatment, rtM204 ＋ rtL180 ＋ rtT184 mutation is common in genotype B and C ETV re-sistance patients.