基于颜色判定的DNA环介导恒温扩增法快速检测金黄色葡萄球菌  

Colorimetric detection of Staphyloccocus aureus by loop-mediated isothermal amplification method

作　　者：李环[1] 刘威[1] 邹大阳[1] 杨展[1] 赵向娜[1] 李雪莲[1] 崔茜[1] 王思淼[1] 魏晓[1] 王雪松[1] 黄留玉[1] 袁静[1] 

机构地区：[1]军事医学科学院疾病预防控制所,北京100071

出　　处：《军事医学》2013年第12期947-950,959,共5页Military Medical Sciences

基　　金：国家"十二五"科技支撑计划资助项目(2011ZX10004-01)

摘　　要：目的建立基于颜色判定的DNA环介导恒温核酸扩增法(loop-mediated isothermal amplification of DNA,LAMP)快速检测金黄色葡萄球菌(SAU)技术。方法利用LAMP法针对SAU特异基因nuc基因设计7套引物,通过对7套引物扩增效率进行比较得到最佳引物组合来快速检测SAU,采用两种结果判断方法在50 min内完成反应。结果 LAMP方法检测核酸的最低浓度为7.16 pg/μl,灵敏度为PCR法的10倍(PCR法为71.6 pg/μl),同时采用22种同源或相似菌种进行特异性检测,结果表明具有良好的特异性。结论 LAMP方法检测过程简单,实验装置简便,反应结果肉眼可视化辨别,灵敏度高,特异性强,特别适合基层防疫与检疫部门对SAU的快速检测。Objective To develop a loop-mediated isothermal amplification （LAMP） assay for Staphyloccocus aureus. Methods Seven sets of primers were designed to recognize S. aureus-specific gene nuc. One set was selected as the best primer, and the reaction was evaluated by two methods in 50 min. Results The sensitivity of LAMP with a detection limit of 7.16 pg/μl was 10 times that of PCR, and the primers designed had a good specificity. Conclusion The LAMP method is not only a potential and valuable means for detection of S. aureus, thanks to its rapidity, simplicity and low cost, but is suitable quick detection.

关 键 词：nuc基因 DNA介导恒温核酸扩增法 金黄色葡萄球菌 快速检测 

分 类 号：R446.5[医药卫生—诊断学] R378.1[医药卫生—临床医学]