利用环介导恒温扩增法快速检测溶组织梭菌  

Rapid detection of Clostridium histolyticum basing on loop-mediated isothermal amplification

作　　者：蒋栋能[1] 刘畅[1] 彭利民[1] 蒲晓允[1] 

机构地区：[1]第三军医大学第二附属医院检验科,重庆400037

出　　处：《国际检验医学杂志》2012年第20期2504-2507,共4页International Journal of Laboratory Medicine

基　　金：重庆市科委重点攻关项目(2011GGB058)

摘　　要：目的利用环介导恒温扩增技术,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(LAMP)方法。检测该LAMP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。(3)检测该LAMP方法对不同浓度溶组织梭菌的扩增情况,观察其最低检测限,对其灵敏度进行评价。结果设计出针对溶组织梭菌的LAMP检测方法。该LAMP检测方法只针对溶组织梭菌有扩增,对其他干扰菌不扩增,显示出良好的特异性。该LAMP检测方法对溶组织梭菌的最低检测限为1×101CFU/mL,显示其灵敏度较高。结论该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 Basing on the loop-mediated isothermal amplification （LAMP）technology,design a rapid detection method of Clostridium histolyticum, studied the reaction characteristics, and to be applied to clinical inspection of C. histolyticum in gas gan- grene. Methods By gene alignment and primer design,a set of LAMP method for detection of C. histolyticum was designed. The LAMP method of detect C. histolyticum and other interference bacteria was tested on their specific evaluation. Detection of the LAMP method with different concentrations of C. histolyticum was amplificated to observe the minimum detection limit,to test its sensitivity. Results We had designed a LAMP method for detection of C. histolytieum. The LAMP method could only amplify C. histolyticum, not amplify the other interference bacteria, showing a good specificity. The LAMP method for detection of C. histolyti eum minimum limit was 1 × 101 CFU / mL, showed a high sensitivity. Conclusion The LAMP method designed for detection of C. histolyticum has a good specificity and high sensitivity,and can be used for detection of C. histolyticum in clinical point-of-care-tes- ting（POCT）.

关 键 词：环介导恒温扩增 溶组织梭菌 快速检验 

分 类 号：R440[医药卫生—诊断学]