环介导等温扩增快速检测人类嗜T淋巴细胞病毒Ⅰ型方法建立  

DEVELOPMENT OF THE LOOP-MEDIATED ISOTHERMAL AMPLIFICATION METHOD FOR RAPID DETECTION OF HUMAN T-CELL LYMPHOTROPIC VIRUS TYPE Ⅰ

作　　者：李涌[1] 白松涛 许欣[1] 刘渠[3] 徐亚军[3] 陈应坚[3] 甘莉萍[3] 杨慧[3] 杨坤祥[3] 

机构地区：[1]四川大学华西公共卫生学院,成都610041 [2]深圳市龙岗区血站 [3]深圳市龙岗区疾病预防控制中心

出　　处：《现代预防医学》2010年第4期728-732,共5页Modern Preventive Medicine

摘　　要：[目的]建立环介导等温扩增(LAMP)快速检测人类嗜T淋巴细胞病毒Ⅰ型方法。[方法]将HTLV-Ⅰ的PX基因转染pUC57构建HTLV-Ⅰ检测参考菌株,针对PX基因设计LAMP和PCR引物,用正交实验设计优化反应体系,并考察方法的特异性和灵敏度。[结果]25μlLAMP最佳反应体系为:外引物0.2μmol/L、内引物1.8μmol/L、dNTP2000μmol/L、Mg2+4mmol/L、甜菜碱0.4mol/L、BstDNA聚合酶8U;最佳反应条件为64℃恒温反应60min。[结论]LAMP方法具有灵敏、特异、操作简便、成本低廉的特点,可为HTLV-I的快速检测提供新手段。[ Objective ] To develop a loop-mediated isothermal amplification method for rapid detection of Human T-cell Lymphotropie Virus Type I. [Methods] HTLV- I reference stain was constructed by transfect a part of PX gene into the pUC57 plasmid, and then LAMP and PCR primers were specially designed to amplify the PX gene. Orthognnal experimental design was adopted in the optimization, also the specificity and sensitivity of the amplification was evaluated. [ Results] The best reaction components concentration was 0.2 μmoL/L out primers, 1.8 μmol/L inner primers, 2000 μmol/L dNTP. 4 mmol/L Mg^2＋, 0.4 moI/L betaine and 8 U BstDNA pelymerase. The most suitable reaction condition was constantly reacting in 64℃ for 60min. [Conclusion] LAMP method established is an effective, high sensitivity, specificity and low-cost method. It could be a new method for rapid detection and identification of HTLV-I.

关 键 词：人类嗜T淋巴细胞病毒I型 环介导等温扩增 

分 类 号：R733.7[医药卫生—肿瘤]