153例慢性HBV感染者核苷和核苷酸类药物的耐药变异位点分析

Drug resistance mutations related to nucleos( t) ide analogues in Chinese patients with chronic HBV infection: an analysis of 153 cases

目的通过对153例慢性HBV感染者进行HBV逆转录酶区扩增测序,了解目前上市的核苷和核苷酸类药物耐药位点检出情况,并分析是否存在与替诺福韦耐药可能相关的耐药位点。方法 153例患者分为2组,其中78例是未使用过核苷和核苷酸类药物治疗的HBV携带者,75例是核苷和核苷酸类药物经治的慢性乙型肝炎患者。通过PCR扩增HBV的逆转录酶(RT)区,进行基因测序分析,了解核苷和核苷酸类药物的耐药位点及替诺福韦相关的耐药位点的检出情况。正态分布资料采用t检验,非正态分布的资料采用秩和检验。结果在未治疗组有7例检出核苷和核苷酸类相关的耐药变异,占该组的8.97%,但并无上述替诺福韦耐药相关的位点。治疗组检出16例耐药变异,占该组的21.33%,其中1例为rtA181T+rtN236T,可能与替诺福韦耐药相关。结论未治疗组中有8.97%的患者检出耐药变异。2组患者中耐药变异的检出率与患者的性别、年龄、病毒基因型均无显著相关。所有患者中仅1例检出rtA181T+rtN236T。提示中国慢性HBV感染者体内预存替诺福韦可能耐药位点的比例很低。

Objective To amplify and determine the hepatitis B virus （HBV） reverse transcriptase （RT） sequences of 153 Chinese patients with chronic HBV infection and investigate the drug resistance mutations related to nucleos（t） ide analogues, and to analyze if there are drug resistance mutations which might lead to tenofovir resistance. Methods A total of 153 patients with chronic HBV infection were divided into two groups： 78 HBV carriers who had never used nueleos（t）ide analogues （NAs） and 75 patients with chronic hepatitis B who had been treated with NAs. Their serum samples were used for HBV DNA extraction and RT gene amplification by PCR, and the RT sequences were analyzed to identify the known drug resistance mutations related to NAs and detect the drug resistance mutations related to tenofovir. The obtained data were statistically analyzed using SPSS 19.0; the Student＇ s t test was used for normally distributed data, and the Wilcoxon＇ s rank sum test for non - normally distributed data. Results Drug resistance mutations related to NAs were detected in 7 cases （8.97%） of the untreated group, but there was no drug resistance mu- tation related to tenofovir. Drug resistance mutations were detected in 16 cases （21.33%） of the treated group, with rtA181T ＋ rtN236T mutations in one case, which might be related to tenofovir resistance. Conclusion The drug resistance mutation detection rate in untreated group was 8. 97%. The drug resistance mutation detection rate was not significantly correlated with HBV genotype and patient＇ s sex and age in either group. The rtA181T ＋ rtN236T mutations were detected in only one case of the treated group, which indicates a very low rate of pre - existence of tenofovir -related drug resistance mutations in Chinese patients with chronic HBV infection.