拉米夫定治疗慢性乙型肝炎中变异病毒株的动态变化  

Dynamics of hepatitis B virus variations during lamivudine treatment

作　　者：田国保[1] 席宏丽[1] 田地[1] 崔建军[1] 于敏[1] 陆海英[2] 曾争[2] 

机构地区：[1]北京大学第一医院感染疾病科病毒研究室,北京市100034 [2]北京大学第一医院感染疾病科,北京市100034

出　　处：《世界华人消化杂志》2007年第30期3245-3250,共6页World Chinese Journal of Digestology

摘　　要：目的:建立定量检测及监测乙肝病毒DNA的rtM204I/V(YMDD)及rtL180M变异的方法.方法:以克隆、测序鉴定及构建乙肝病毒DNA YMDD及变异(YIDD和YVDD)质粒作为标准,4例接受拉米夫定治疗(100 mg/d,疗程48 wk以上)的慢性乙型肝炎患者血清标本为对象,用焦磷酸测序的方法检测变异位点核苷酸的频率,并计算变异株的含量比例,通过检测不同变异株类型的标准质粒,确定测序峰图的背景信号.结果:通过检测标准质粒后,确定背景信号为5%,变异位点调整后的核苷酸频率转化为变异病毒株的比例后,4例患者治疗前均检测出变异的病毒株(4.5%-33%),并且随治疗时间延长呈增多的趋势.病毒载量及ALT分析提示基因型耐药发生后,将发生病毒学反跳及临床耐药.结论:焦磷酸测序可以定量检测及动态监测变异病毒株的含量.拉米夫定耐药突变株在拉米夫定治疗前已存在,并随治疗时间的增加而增长,出现病毒学反弹及临床耐药.AIM： To use pyrosequencing to establish a method for monitoring hepatitis B virus mutants resistant to lamivudine, METHODS： Plasmids cloned, sequenced and constructed from the lamivudine-resistant HBV mutants at rtM204I/V （YMDD motif） and rtL180M were used as a standard, The frequencies of mutation-site nucleotides were detected by pyrosequencing in sera from 4 patients who underwent lamivudine therapy. HBV DNA with rtM204I, rtM204V and wildtype plasmids were constructed for subtracting the background signal frequencies.was monitored for 4 to 8 months along with the gradually increasing HBV variations at positions of rtM204I/V and rtL180M. CONCLUSION： Pyrosequencing is an effective method to quantify and monitor HBV variations during lamivudine therapy. HBV lamivudineresistant mutants exist before lamivudine treatment and gradually increase during treatment, and then virological breakthrough and clinical resistance occur.

关 键 词：焦磷酸测序 拉米夫定 治疗 慢性乙型肝炎 变异病毒株 动态变化 

分 类 号：R512.6[医药卫生—内科学]