环介导等温扩增快速可视化检测B族链球菌的应用研究  

A loop-mediated isothermal amplification assay for the rapid visualization detection of group B streptococcus

作　　者：王行[1] 汪骅[2] 张立 WANG Xing;WANG Hua;ZHANG Li(Department of Clinical Laboratory Putuo District People＇s Hospital ,Shanghai 200060,China;Department of Clinical Laboratory ,Renji Hospital ,Shanghai Jiaotong University School of Medicine ,Shanghai 200120,China)

机构地区：[1]上海市普陀区人民医院检验科,上海200060 [2]上海交通大学医学院附属仁济医院检验科,上海200120

出　　处：《国际检验医学杂志》2018年第19期2366-2368,2372,共4页International Journal of Laboratory Medicine

基　　金：上海市普陀区人民医院院级课题(2015Rm101)

摘　　要：目的建立环介导等温扩增(LAMP)快速检测B族链球菌(GBS)的方法并探讨此方法在临床诊断中的应用价值。方法培养GBS,提取病原体DNA,设计GBS 16S-23SrRNA基因间隔区LAMP引物,优化并建立LAMP检测GBS的方法。分析LAMP法检测GBS DNA灵敏度,并与聚合酶链反应(PCR)法进行对比。分析LAMP法检测GBS DNA的特异度。结果 LAMP法可在1h内实现GBS DNA的可视化检测,检测限为20fg,灵敏度高于PCR法(200fg)。LAMP法检测50例经PCR法确诊的GBS感染的临床样本,结果均为阳性,凝固酶阴性葡萄球菌、大肠埃希菌、淋病奈瑟菌、白色念珠菌的检测结果均为阴性。结论 LAMP检测GBS快速、简便、灵敏高且特异度强,检测不需任何设备,借助荧光笔肉眼直接判读,有望成为社区及基层医院广泛开展的新方法。Objective To establish a loop-mediated isothermal amplification （LAMP） assay for the rapid visualization detection of group B streptococcus and application prospect of this method was discussed. Meth- ods Culture of B-group streptococcus and extraction of pathogen DNA. With the 16S-23S rRNA intergenic spacers as targets, four sets of LAMP primers were designed for identification group B streptococcus. Group B streptococcus DNA was used to determine the sensitivity and specificity of this method compared to PCR method. Results GBS DNA amplification was detected within lb. The detection limit of LAMP method was 20 fg DNA template per reaction. The sensitivity of LAMP was 10 times compared with conventional PCR. The lMAP assays were then validated by analysis with samples from 50 GBS patients who had previously been analyzed via PCR. The results from the LMAP assay were in perfect agreement with PCR method. Con- clusion LAMP assay can be used as a sensitive,rapid,and simple detection tool for the visualization detection of group B streptococcus. The methods do not require any additional user interaction is ideal for use of portable microfluidic platforms doctor′s offices in the field or at the bed-side.

关 键 词：环介导等温扩增 可视化检测 B族链球菌 

分 类 号：R440[医药卫生—诊断学]