未治和经治慢性乙型肝炎患者的病毒基因型分析及其核苷酸类似物耐药突变的模式特征

Detection of hepatitis B virus genotyping in untreated or treated patients of chronic hepatitis B and its mutation patterns of the nucleotide analogues-resistant mutants

目的检测分析未治和经治慢性乙型肝炎患者病毒基因型及其核酸类似物耐药突变位点的模式特征,为慢性乙型肝炎患者的抗病毒治疗方案提供依据。方法随机选取74例未治和66例经治的慢性乙型肝炎患者,PCR反向斑点杂交法检测其血清乙型肝炎病毒的基因型及核苷酸类似物耐药基因突变位点。结果未治患者与经治患者的血清乙型肝炎病毒基因型均以B型为主,两组差异无统计学意义(P>0.05),未治患者HBV C型的比例低于经治患者(P<0.05),B+C和B+D型的比例则高于经治患者(P<0.05)。未治患者同样存在耐药基因突变:尽管其B型和C型HBV的耐药突变比例均低于经治患者(P<0.05),但其B+D型HBV的耐药突变比例高于经治患者(P<0.05),且以236T单基因突变为主;经治组患者HBV耐药突变单位点180M、204V和204I例数均高于未治组,而且经治组患者比未治组患者有更多的多位点耐药突变(P<0.05)。结论 HBV B基因型是本研究主要的基因型。未治患者亦存在耐药基因耐药突变,以B+D型HBV的耐药基因突变和236T单基因突变为特征;与未治患者相比,经治患者有更多的单基因位点和多基因位点的突变。未治和经治慢性乙型肝炎患者,均应在治疗过程中定期监测NAs耐药突变发生,以便及时选择最佳治疗方案。

Objective To investigate and identify prevalent genotypes of hepatitis B virus（HBV）among untreated or treated patients with chronic hepatitis B（CHB） and the mutation pattern of the nucleotide analogues-resistant mutants. Methods A total of 74 cases with CHB who did not receive the treatment of lamivudine and any other antivirus drugs and 66 cases with CHB who receive the treatment of lamivudine and any other antivirus drugs were randomly chosen. After sample collection and HBV DNA extraction, PCR reverse spot hybridization was used to detect HBV genotypes and the nucleotide analogues（NAs）-resistant mutants. Results The main Genotypes of HBV is the B type in both untreated and treated patients. The rate of HBV C in the untreated patients was less than that of the treated patients（P〈0.05）, while B＋C and B＋D were higher in the untreated patients compared with the treated patients（P〈0.05）. The NAs-resistant mutants were found in both untreated and treated patients of CHB. Despite of the less rate of the NAs-resistant mutants in the virus genotypes B ＋ C and higher rate of that in the virus genotypes B ＋ D among untreated patients compared with treated patients（P〈0.05）, the single locus of NAs-resistant mutants were revealed 236 T as the main mutation pattern in the untreated patients（P〈0.05）. While the 180 M, 204 V and 204 I as the main single locus mutation pattern in the treated patients（P〈0.05）and, more multiple gene loci mutation were detected among the treated patients compared with untreated patients. Conclusion HBV B is the main genotypes in this research. NAs-resistant mutants existed in the untreated patients. The mutant sites in the treated patients were more both in single gene locus and multiple gene loci than that in the untreated patients. Investigation of NAs-resistant mutants before treatment is crucial for the success of the treatment. In whole course of antiviral treatment, NAs-resistant mutants should be monitored for the optimal schedule.