利用环介导恒温扩增法快速检测伤寒杆菌  

Rapid detection of Typhoid Bacilus by loop-mediated isothemal amplification assay

作　　者：刘琳琳[1] 蒋栋能[1] 蒲晓允[1] 

机构地区：[1]第三军医大学新桥医院检验科,重庆400037

出　　处：《国际检验医学杂志》2013年第5期534-535,538,共3页International Journal of Laboratory Medicine

基　　金：重庆市科委重点攻关项目(2011AB5035)

摘　　要：目的利用环介导恒温扩增技术(LAMP),设计快速检测伤寒杆菌的方法,研究其反应特性,以期能应用于临床现场检验。方法 (1)通过基因比对与引物设计,建立针对伤寒杆菌的LAMP检测方法。(2)根据该方法对伤寒杆菌及其他干扰菌进行检测时的扩增情况,对其特异性进行评价。(3)根据该LAMP方法对不同浓度伤寒杆菌的扩增情况,得出其最低检测限,对其灵敏度进行评价。结果该LAMP检测方法只针对伤寒杆菌有扩增,对其他干扰菌不扩增,显示出良好的特异性。该方法对伤寒杆菌的最低检测限为1×101CFU/mL,其灵敏度较高。结论本试验设计出了针对伤寒杆菌的LAMP检测方法,该LAMP检测方法具有良好的特异性和较高的灵敏度,能够用于伤寒杆菌的临床快速检测。Objective Use loop mediated isothermal amplification（LAMP）technology to design method for rapid Salmonella typhi detection,and study the reaction＇s characteristics in order to apply to the clinical inspection.Methods（1）By gene alignment and primer design,build LAMP detection method for Salmonella typhi.（2）According to the amplification by LAMP method for the detection of Salmonella typhi and other interference bacteria,its specific assessment was made.（3）Detection of the LAMP method for different concentrations of Salmonella typhi amplification,observe the minimum detection limit,and sensitivity assessment was made.Results The LAMP method only worked for Salmonella typhi,other interference bacteria were not amplificated,showing good specificity.The minimum detection limit of the method of Salmonella typhi was 1×101 CFU/mL,the sensitivity was good.Conclusion The research designed a LAMP method for Salmonella typhi detection,the LAMP method has good specificity and high sensitivity,and can be used for Salmonella typhi＇s clinical inspection.

关 键 词：敏感性与特异性 环介导恒温扩增 伤寒杆菌 快速检测 

分 类 号：R446.5[医药卫生—诊断学]