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目的分析慢性乙型肝炎患者HBV逆转录酶区耐药变异情况及rtA181变异准种的分布。方法提取患者血清中HBVDNA,PCR扩增逆转录酶区域,产物测序后经软件比对分析耐药变异情况和基因型;对部分rtA181变异标本进行克隆后测序分析准种分布。结果 489例标本中,检出明确耐药变异265例,在B、C基因型中分布比例存在差异(56.6%vs43.0%,P=0.022)。拉米夫定相关耐药138例(52.1%),以M204I、M204I/V+L180M±L80I/V变异为主;阿德福韦相关耐药35例(13.2%),以N236T+A181T/V较为多见;拉米夫定+阿德福韦相关耐药70例(26.4%),几乎都和A181有关。rtA181准种分析发现1例位于同一病毒株的多耐药组合,且未发现单一A181T变异的病毒准种。结论 HBV耐药变异主要表现为M204和A181相关变异,耐药模式复杂;检测HBV逆转录酶区的变异有助于临床及时发现和确认耐药情况,指导临床合理进行抗病毒用药。
Objective To analyze the variation of drug resistance in HBV reverse transcriptase region in patients with chronic hepatitis B and the distribution of rtA181 variant. Methods HBVDNA was extracted from serum of patients and the region of reverse transcriptase was amplified by PCR. The products were sequenced and analyzed by software to analyze the variation of drug resistance and genotype. The partial rtA181 samples were cloned and sequenced to analyze the quasispecies distribution. Results In 489 specimens, 265 cases with clear resistance mutations were detected. There were differences in the distribution of B and C genotypes (56.6% vs 43.0%, P = 0.022). There were 138 cases (52.1%) with lamivudine related resistance, with M204I, M204I / V + L180M ± L80I / V mutation as the main drug resistance. Adefovir resistance was related to 35 cases (13.2%) with N236T + A181T / V More common; lamivudine + adefovir-related 70 cases of drug resistance (26.4%), almost all related to A181. Quasi-species analysis of rtA181 revealed 1 multi-drug resistant combination located in the same virus strain, and no single virus variant of A181T was found. Conclusions HBV resistance variation is mainly related to the mutation of M204 and A181, and the pattern of drug resistance is complex. Detecting the variation of HBV reverse transcriptase region is helpful to find out and confirm the drug resistance clinically and guide the rational use of antiviral drugs.