340例慢性乙型肝炎患者乙型肝炎病毒多位点耐药相关突变分析  

Multiple-site analysis of HBV drug-resistant mutations in 340 patients with chronic hepatitis B

作　　者：徐东平[1] 刘妍[1] 成军[2] 李晓东[1] 戴久增[1] 李乐[1] 梁兆玲[1] 白丽[1] 钟彦伟[1] 许智慧[1] 任晓强[1] 张玲霞[1] 

机构地区：[1]解放军第三0二医院传染病研究所病毒性肝炎研究室,北京100039 [2]北京地坛医院

出　　处：《中华肝脏病杂志》2008年第10期735-738,共4页Chinese Journal of Hepatology

基　　金：国家重点基础研究发展计划（“973”）项目（2007CB512803）;北京市科技计划课题（Z07050700690702）

摘　　要：目的分析慢性乙型肝炎患者HBV逆转录酶基因与核苷（酸）类似物耐药相关的12个位点上的突变情况及其临床意义。方法提取血清HBVDNA，扩增HBV逆转录酶基因，对PCR产物进行DNA双向测序，对测序成功的样本进行基因型分析。检测逆转录酶基因12个位点上的碱基突变情况，分析不同核苷（酸）．类似物使用情况、患者的耐药相关突变情况及不同核苷（酸）类似物耐药的突变形式。结果检出拉米夫定耐药突变63例，阿德福韦耐药突变10例，恩替卡韦耐药突变8例，替比夫定耐药突变l例。拉米夫定耐药突变中以M204V和M204I最常见，前者通常伴随L180M突变，后者常单独出现；阿德福韦耐药中以N236T±A181位碱基替换为主；恩替卡韦耐药突变发生在拉米夫定耐药基础上，以T184位碱基替换为主；替比夫定的耐药突变为M204I。少数未接受过核苷（酸）类似物治疗的患者也可检出耐药相关突变。结论检测HBV逆转录酶基因多位点耐药相关突变，有助于临床及时发现和确认乙型肝炎患者是否存在HBV耐药，合理进行抗病毒治疗。Objective To analyze HBV drug-resistant mutations against nucleos（t）ide analogues at 12 reported sites in 340 patients with chronic hepatitis B. Methods Serum HBV DNA was extracted and a nested PCR assay was employed for the reverse transcriptase （RT） gene amplification. Direct sequencing of PCR product was performed. The significance of detected mutations was analyzed in view of clinical data of the patients. Results Drug-resistant mutations were detected in 68 patients taking lamivudine （LAM）, I0 taking adefovir （ADV）, 8 taking entecavir, and 1 taking telbivudine （LdT）. M204V and M204I were the most common LAM-resistant mutations. The former usually emerged with L180M while the latter often emerged alone. N236T ± A181 substitution was the most frequently seen ADV-resistant mutation. ETV-resistant mu- tations occurred on the basis of LAM-resistant mutations and T184 change was the most common form. LdT- resistance was observed as M204I. Interestingly, these drug-resistant mutations were detected in a few pa- tients who had not been treated with nucleos（t）ide analogues. Conclusion Detection of HBV drug-resistant mutations at multiple sites of the viral RT gene is valuable for discovering and verifying drug resistance and thus is very helpful in planning anti-HBV therapy.

关 键 词：肝炎病毒 乙型 突变 耐药 

分 类 号：R512.62[医药卫生—内科学] R373.1[医药卫生—临床医学]