利用环介导恒温扩增法快速检测溶组织梭菌  

Rapid Detection of Clostridium Histolyticum Based on Loop-mediated Isothermal Amplification

作　　者：舒利琼[1] 刘畅[1] 彭利民[1] 蒋栋能[1] 

机构地区：[1]第三军医大学附属新桥医院检验科,重庆400037

出　　处：《现代生物医学进展》2013年第8期1442-1445,共4页Progress in Modern Biomedicine

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),设计快速检测溶组织梭菌的方法,研究其反应特性,以期能应用于气性坏疽溶组织梭菌感染的临床现场检验。方法:通过基因比对与引物设计,设计针对溶组织梭菌的环介导恒温扩增检测方法。然后检测该方法对溶组织梭菌及其他干扰菌的扩增情况,对其特异性进行评价。检测该方法对不同浓度溶组织梭菌的扩增情况,对其灵敏度进行评价。最后与全自动细菌培养法进行比较,评价其与常规检测方法的可比性。结果:本LAMP检测方法只针对溶组织梭菌有扩增,对其他15种菌不扩增,显示出良好的特异性。对溶组织梭菌的最低检测限为1×101CFU/ml,显示其灵敏度较高。与全自动细菌培养法相比,有良好的可比性(P<0.05),很高的灵敏度(92.6%)、特异性(98.1%)及准确度(97.3%)。结论:本试验设计出了针对溶组织梭菌的LAMP检测方法,该LAMP检测方法具有良好的特异性较高的灵敏度,能够用于溶组织梭菌的临床现场检验。Objective： Based on the loop-mediated isothermal amplification （LAMP）technology, we designed a rapid detection method ofC. histolyticum, studied the reaction characteristics, and to be applied to clinical inspection ofC. histolyticum in gas gangrene. Methods： By gene alignment and primer design, a LAMP method ofC. histolyticum was designed. The method was tested on its specific evaluation by detecting C. histolyticum and other interference bacteria. Then different concentrations of C. histolyticum was amplificated to test its sensitivity. Finally, it was compared with the automatic bacterial culture method to evaluate the comparability to the conventional method. Results： A LAMP method ofC. histolyticum was designed. The LAMP method could only amplify C. histolyticum, not amplify the other 15 bacteria, showing a good specificity. Its minimum limit was 1 × 10ICFU / ml, showed a high sensitivity. Compared with the automated bacterial culture method, it has a good comparability （P〈0.05）, high sensitivity （92.6%）, specificity （98.1%） and accuracy （97.3%）. Conclusion： The LAMP method of C. histolyticum has a good specificity and high sensitivity, and can be used for detection of C. histolyticum in clinical piont-of-care-testing （POCT）.

关 键 词：环介导恒温扩增 溶组织梭菌 临床现场检验 

分 类 号：R446[医药卫生—诊断学] R373[医药卫生—临床医学]