环介导等温扩增技术在钩端螺旋体病监测中的应用

The application of loop mediated isothermal amplification in surveillance for leptospira interrogans

目的:评价LAMP技术在钩端螺旋体检测与监测中的可行性。方法:采用LAMP技术对感染病人血清及钩体标准菌株感染鼠肾进行检测,同时与卫生行业标准推荐的引物G1/G2 PCR方法进行对比。结果:LAMP快速检测方法 60 min内即可完成检测,具有较高的检测灵敏度,对纯培养的钩体检测灵敏度可达1.5 CFU/ml,比PCR法高出两个数量级,结果肉眼直接可观。同时对21种共42株细菌进行LAMP扩增,仅钩体结果为阳性,显示该LAMP方法具有较强特异性。结论:LAMP方法操作简便、特异性强、灵敏度高且成本低廉,在钩体的快速检测和疫情监控中具有良好的应用前景。

Objective：To discuss the feasibility of LAMP for detecting and monitoring Leptospira interrogans.Methods： The specificity and sensitivity of detection for leptospiral DNA in human serum and rat kidney based on Loop mediated isothermal amplification（LAMP） were compared with those of the recommended standard G1/G2 PCR.Results： 42 bacterial strains were selected to test specificity of the LAMP method and only Leptospira interrogans showed positive result by LAMP,indicating the high specificity of the LAMP method.The LAMP method could be completed in 60 min and showed a good sensitivity of 1.5 CFU/ml for cultured Leptospira interrogans.Conclusion： The LAMP assay is a sensitive,rapid and simple tool for the detection of Leptospira interrogans,performing a bright future for the detection and monitoring of Leptospira interrogans.