拉米夫定耐药HBV毒株RT区变异分析  

The mutations analysis of the reverse transcriptase domain of HBV polymerase in Lamivudine-resistant patients

作　　者：赵卫峰[1] 邵幼林[1] 甘建和[1] 陈良云[1] 朱义玲[1] 毛日成[2] 张继明[2] 

机构地区：[1]苏州大学附属第一医院感染科,江苏苏州215006 [2]复旦大学附属华山医院感染科,上海200040

出　　处：《中国实用内科杂志》2008年第2期119-121,共3页Chinese Journal of Practical Internal Medicine

基　　金：江苏省卫生厅科研项目(H200711);上海市科委临床医学重点项目(054119529)

摘　　要：目的研究拉米夫定(LAM)治疗后出现HBV病毒学突破患者HBVRT区变异位点和变异类型。方法研究对象选自2004年4月至2007年3月在苏州大学附属第一医院门诊或住院治疗的慢性乙型肝炎患者,用聚合酶链式反应(PCR)方法扩增LAM治疗后出现HBV病毒学突破患者的血清HBVRT区基因,对PCR产物直接测序,用Chromas2.0软件分析HBVRT区基因的核苷酸和氨基酸差异、变异类型。结果109例患者在拉米夫定耐药后出现病毒学突破,其中94例出现拉米夫定耐药相关性变异,包括YMDD变异93例,单独rtA181T变异1例。13例(11.93%)患者经测序分析未发现YMDD变异,但用限制性内切酶片段长度多态性(RFLP)方法检测,均发现有YMDD变异。测序结果发生变异位点和出现频率:rtM204V/I93例(85.3%)、rtL180M51例(46.9%)、rtV173L/M7例(6.4%)、rtV207M/L/I4例(3.7%)、rtA181T4例(3.7%)、rtT184I/S/M2例(1.8%)、rtM250L2例(1.8%)。变异类型:rtM204V/I、rtA181T、rtM204V/I+rtL180M、rtM204V/I+rtL180M+rtV173M等。结论拉米夫定耐药主要变异类型为rtM204V/I变异,常伴随rtL180M和rtV173L/M变异;少数拉米夫定耐药患者在阿德福韦和恩替卡韦治疗前即已产生阿德福韦和恩替卡韦耐药相关性变异。Objective To study the nucleotide substitution sites and patterns of the reverse transcriptase （RT） domain of HBV polymerase of patients with viral DNA breakthrough under LAM therapy. Methods Between April 2004 and March 2007, the subjuts were selected from patients with chronic hepatitis B who were treated in the First Affiliated Hospital of Soochow University. HBV strains were amplified by PCR using specific primers flanking B-C regions of RT and S gene. The PCR products were directly analyzed by sequencing. The difference of nucleotide, amino acid and mutation patterns of RT region was classified by using Chromas2. 0 software. Genetyping HBV was analyzed by using Clustalxl. 81. msw. Resuits Totally 93 YMDD mutations and 1 patient（ rtA181T）occurred in 109 patients experiencing virologic breakthrough. The 13 patients （8.6 % , 13/109）were not found YMDD mutation by sequencing but detected by RFLP. The substitutions at residues of LAM-r were rtM204V/I （ n = 93,85.3% ）, rtL180M （ n = 51,46. 9% ） , rtV173L/M （ n = 7,6.4% ）, rtV207M/L/l（n =4,3.7% ） ,rtAl81T（n =4,3.7% ） ,and rtM250L（ n = 2,1.8% ）. The major patterns of LAM-r HBV were identified, namely, rtM204V/I, rtA 181 T, rtM204V/I ＋ rtL180M, rtM204V/I ＋ rt L180M ＋ rtV173 M and so on. Conclusion The major mutations with LAM-r are rtM204V/I and usually companied by rtL180M and rtVI73L/M mutations. A few LAM-r patients have adefovir and entecavir resistance before switihing to adefovir and entecavir.

关 键 词：肝炎 乙型 慢性 肝炎病毒 变异 拉米夫定 耐药 

分 类 号：R5[医药卫生—内科学]