多重环介导核酸等温扩增技术检测血液中常见病原体  

Development of Multiplex Loop Mediated Isothermal Amplification Method for the Simultaneous Detection of HBV,HCV,HIV and TP

作　　者：梁超[1] 成思佳[1] 王聪[1] 初亚男[1] 奚涛[1] 周国华[1,2,3] 

机构地区：[1]中国药科大学生命科学与技术学院,江苏南京210009 [2]华东医学生物技术研究所,江苏南京210002 [3]南京军区南京总医院药理科,江苏南京210002

出　　处：《现代生物医学进展》2013年第1期141-145,77,共6页Progress in Modern Biomedicine

基　　金：江苏省社会发展项目(BE2010605;BE2011607)

摘　　要：背景:血液安全性筛查是输血前必要检测项目。目前临床采用血清学检测技术,存在较长的检测窗口期,易产生假阴性检测结果,造成输血交叉感染。目的:建立多重环介导核酸等温扩增技术,实现在一管反应体系内同时检测四种病原体:乙肝病毒,丙肝病毒,艾滋病毒和梅毒螺旋体。方法:通过限制性酶切处理多重环介导核酸等温扩增产物,利用酶切产物的长度分析扩增产物的种类,从而分析待测样本中含有何种血液病原体。结果:检测164例临床样本,其检测结果可以通过琼脂糖电泳,聚丙烯酰胺凝胶电泳及芯片电泳分析,且均可实现对多重扩增产物的酶切片段进行区分和鉴别。结论:多重环介导核酸等温扩增技术可以同时单管检测多种待测血液病原体,可以为临床提高简单、快速、高灵敏和高特异的检测技术。The blood safety screening was an essential test before blood transferring. The window period of serologic test was long which may lead to false negative results and cause infection of blood pathogens. Objective： In order to achieve screening of four blood pathogens, HBV, HCV, HIV and TP in a single reaction, we planned to establish a novel method based on multiplex LAMP reaction. Methods： In this study, we designed four sets of the LAMP primers for conservative sequences of HBV, HCV, HIV and TP, which constructed a multiplex LAMP （mLAMP） method by combining these four sets totaling 32 primers. The mLAMP method was distinguishable among HBV, HCV, HIV and TP, simultaneously, due to the subsequent restriction enzyme analysis. Results： Of 164 blood samples collected from China, which were analyzed by agarose electrophoresis, PAGE and microchip electrophoresis. The detection results were same as serologic test. Conclusions： These findings indicate that this mLAMP method is a new convenient tool for simultaneous detection of the pathogens in blood.

关 键 词：多重环介导核酸等温扩增技术 限制性酶切分析 血液病原体检测 

分 类 号：R446.11[医药卫生—诊断学]