740例HBV感染者血清HBV逆转录酶区耐药位点分析

Mutation of hepatitis B virus DNA reverse transcriptase in 740 patients with hepatitis B and liver cirrhosis

目的：了解乙型肝炎病毒（HBV）感染者 HBV 基因型的构成状况及 HBV P 区基因变异的特点。方法对2010年8月～2011年10月送检的740份 HBV 感染者血清，采用 ABI 基因测序仪检测 HBV 耐药位点和基因型，应用 SPSS15.0软件进行统计分析。结果在740例 HBV 感染者中，检出 HBV B 基因型40例（5.41%），C 基因型695例（93.92%），D 基因型5例（0.68%）；与拉米夫定、阿德福韦酯和恩替卡韦耐药明确相关的位点突变377例（50.95%），其中195例（51.72%）患者既往有明确的应用核苷（酸）类似物（NA）史，在这195例发生耐药的患者中，B基因型12例，C 基因型183例，两基因型间耐药发生率无明显差异，他们中包括 CHB 患者118例，肝硬化患者77例，主要耐药变异模式为 M204V＋L180M、M204I、M204I＋L180M 和 A181V，两组间 ALT、HBV DNA 载量和主要耐药变异模式检出率均无统计学差异；HBV DNA 载量是影响 NA 耐药的相关因素（x2=0.496，P〈0.001），但耐药与年龄、性别、疾病严重程度（CHB 或肝硬化）和 ALT 水平无显著性相关；此外，本文还检出多处未知变异位点，如 rt64、rt126、rt178和 rt129等。结论核苷（酸）类似物的使用将伴随病毒变异的发生，其耐药基因变异位点具有一定的特点及规律性，动态监测 HBV DNA 载量对早期发现耐药有一定的价值。

Objective Hepatitis B virus （HBV） genotypes and pre-core mutations were studied in patients with HBV infection. Methods Viral DNA was extracted from serum samples of 740 patients with hepatitis B and liver cirrhosis from August 2010 to October 2011,and HBV genotypes and pre-core mutations were detected by ABI sequencing instrument. Results Among 740 patients with HBV infection,40（5.41%） were with HBV genotype B,695（93.92%） were with genotype C,and 5（0.68%） were with genotype D infection;377（50.95%） patients car-ried mutations related to lamivudine,adefovir dipivoxil and entecavir resistance,among whom 195 patients（51.72%） has a definite history of application of nucleos（t）ide analog （NA）. Out of the 195 patients,there were 12 patients with HBV genotype B and 183 with genotype C infection. Also among these patients with definite NA history,118 patients were with CHB,and 77 patients were with liver cirrhosis,and the major resistant mutations were M204V＋L180M,M204,M204I＋L180M and A181V;Serum HBV DNA load was related to NA resistance（x2=0.496, P〈0.001）,but not age,gender,disease severity （CHB or cirrhosis） and ALT levels;Besides,a number of new muta-tions,e.g. rt64,rt126,rt178,rt129 and so on were found in this series. Conclusions The administration of NA might results in mutation of hepatitis B virus and drug resistance,and the mutations have certain characteristics. Dynamic monitoring of serum HBV DNA load will help early detection of gene mutation and drug resistance.