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目的研究乙型肝炎病毒(HBV)耐药相关位点的变异模式与HBV基因型的相关性。方法收集3 255例有核苷(酸)类似物应用史,并且HBV DNA高于检测水平或者出现病毒学反弹的慢性乙型肝炎患者血清,提取HBV DNA,用PCR扩增逆转录酶基因,产物直接进行DNA测序。结果 3 255例样本中,C基因型2 378例,B基因型858例,D基因型19例。1 671例检测到耐药相关基因型(1 671/3 255,51.3%;包括1 126例M204变异,297例A181变异,50例N236变异,198例M204/A181/N236组合变异)。其中C基因型耐药1 317例(1 317/2 378,55.4%),B基因型耐药348例(348/858,40.6%),D基因型耐药6例(6/19,31.6%)。C基因型检测到耐药相关变异的频率高于B基因型(55.4%vs 40.6%,P<0.000 1)。1 523例检测到次级耐药位点及补偿耐药位点(包括rt173、rt180、rt207、rt213、rt214、rt215、rt237、rt238、rt184、rt202、rt250)变异。M204V变异在C基因型中较B基因型多见(15.1%vs12.2%,P=0.043 4)。M204I+L180M变异发生率在C基因型中比B基因型高(8.92%vs 3.26%,P<0.000 1)。N236T变异在B基因型中多见(4.20%vs 0.84%,P<0.000 1),而A181T/V变异在C基因中多见(6.48%vs 1.75%,P<0.000 1)。M204V比M204I易合并L180M代偿性变异及继发恩替卡韦耐药(M204V/I+L180M+T184/S202/M250)(P<0.000 1)。结论 HBV耐药相关位点的变异模式与HBV基因型有一定相关性,对优化药物选择及临床结局有潜在影响,但其机制有待进一步研究。
Objective To study the relationship between HBV genotypes and the variation patterns of hepatitis B virus (HBV) resistance-related loci. Methods A total of 3 255 patients with history of application of nucleoside (acid) analogues were collected, and HBV DNA was detected in serum of patients with chronic hepatitis B whose level of HBV DNA was higher than the detection level or appeared virological rebound. HBV DNA was extracted and the reverse transcriptase gene was amplified by PCR. Direct DNA sequencing. Results Among 3 255 cases, there were 2 378 C genotypes, 858 B genotypes and 19 D genotypes. A total of 1 671 cases were detected with resistance-related genotypes (1 671/3 255 and 51.3%; including 1 126 M204 mutations, 297 A181 mutations, 50 N236 mutations and 198 M204 / A181 / N236 combined mutations). There were 1317 cases (1 317/2 378, 55.4%) of the C genotypes, 348 cases (348/858,40.6%) of the B genotypes and 6 cases (6/19%) of the D genotypes ). The frequency of resistance-related mutations detected in genotype C was higher than in genotype B (55.4% vs 40.6%, P <0.0001). A total of 1 523 cases were detected with secondary resistance and drug-resistant sites (including rt173, rt180, rt207, rt213, rt214, rt215, rt237, rt238, rt184, rt202, rt250). The M204V mutation was more common in the C genotype than the B genotype (15.1% vs 12.2%, P = 0.043 4). The incidence of M204I + L180M mutation was higher in genotype C than genotype B (8.92% vs 3.26%, P <0.0001). N236T mutation was more common in B genotype (4.20% vs 0.84%, P <0.0001), while A181T / V mutation was more common in C gene (6.48% vs 1.75%, P <0.0001). M204V was more susceptible to compensatory mutation L180M than M204I and secondary to entecavir resistance (M204V / I + L180M + T184 / S202 / M250) (P <0.0001). Conclusion The pattern of variation of HBV resistance-related loci has a certain relationship with HBV genotype, which may have potential impact on the choice of drug and clinical outcome. However, its mechanism remains to be further studied.