一种新的HBV表型耐药研究方法的建立  

A new strategy for studying the drug susceptibility of human hepatitis B virus in vitro

作　　者：朱华[1] 苏何玲[1] 刘永明[1] 刘青波[1] 马义丽[1] 许智慧[2] 钟彦伟[2] 徐东平[2] 

机构地区：[1]桂林医学院生物技术学院,541004 [2]解放军第三0二医院肝衰竭诊疗研究中心病毒性肝炎研究室,北京市100039

出　　处：《实用医学杂志》2013年第7期1037-1040,共4页The Journal of Practical Medicine

基　　金：国家“十一五”传染病重大专项子课题(编号:2008ZX10002-011);北京市自然科学基金课题(编号:7091006)

摘　　要：目的:探索建立一种快速、简便、经济的HBV表型耐药分析方法。方法:将野生型pTriEx-HBV1.1表达载体转染HepG2细胞,5h后给予不同浓度的LAM、ADV及ETV处理,隔天换药1次,换药2d后收集培养上清及细胞,用Dnase处理后,real-timePCR检测HBVDNA的水平。结果:上清及细胞内核心颗粒HBVDNA的水平下降2～3个数量级,最大抑制率达98%,EC50值接近。结论:以上清病毒定量代替核心颗粒,建立了基于real-timePCR的HBV表型耐药分析方法,有利于HBV临床治疗的耐药监测。Objective To develop an efficient, simple and economic method for the phenotypic assay of HBV drug susceptibility in vitro. Methods wt PTriEx-HBVI.1 vector was transfected into HepG2 cells. The cells were treated with serial concentration of LAM, ADV and ETV five hours after transfection and renewed on the next day. Supernatant and culture cells were treated with Dnase two days after drug renewing, and the viral DNA was measured by real-time PCR. Results Viral DNA level in superuatant and culture cells decreased by 2~3 log with the maximum inhibition rate of 98%. Conclusion We have developed a new phenotypic assay method based on the real-time PCR, using the supernatant viral quantity instead of intracellular HBV DNA determination.

关 键 词：肝炎病毒 乙型 表型耐药分析 REAL-TIME PCR 

分 类 号：R446.5[医药卫生—诊断学]