Loop mediated isothermal amplification(LAMP) method for detection of Vibrio cholerae

Objective To apply the loop mediated isotheun al amplification (LAMP) method for rapid detection of Vibrio cholerae.Methods A set of four prin ers(two outer and two inner) was designed specifically to recognize the house-keeping genes (mdh) of Vibrio cholerar.The method of LAMP for rapid detection of Vibrio cholerae in food sample was established Cenom is DNAs from 42 strains of 17 hinds of bacteria were amplified with LAMP.The sensitivity,specificity and precision of the method were evaluated.Results Tw enty-one strains of Vibrio cholerae were all found to be positive by the LAMP and no amplicon was observed in other 21 bacterial strains.The most optin al reaction tin a and temperature of the LAMP assay for the mdh gene were 60 min and 65℃,respectively.The detection lin it of the LAMP assay was around 40 fg of Vibrio cholerae genom is DNA and 50 colonies founing units for pure cultrues In addition,the method was applied to detect artificially contaminated food samples,and the detection lin it was 70 cfu/g for non-cultured artificially contaminated.food samples.Conclusion The results suggest that detection of Vibrio cholerae by LAMP is an effective and low-cost procedure with high specificity and sensitivity that requires no specialized equipment.So it can be extensively used for fast screen of samples.