Development of loop-mediated isothermal amplification method for detection of Yersinia pestis

Objective:To develop a method of 1oop-mediated isothermal amplification(LAMP) for detection of Yersinia pestis.Methods: According to conserved F1 gene of Yersinia pestis and principle of LAMP,a set of LAMP reaction system containing four primers were designed.Strains of Yersinia pestis and non-Yersinia pestis were detected with LAMP to evaluate the specificity of the method.The dilution of the Yersinia pestis was detected with LAMP and PCR to evaluate the sensitivity of the method.Results: 10 non-Yersinia pestis strains were found all negative by LAMP,performing high specificity of this method.The detection limit of this LAMP assay was about 20 yersinia pestis strains per reaction.The sensitivity of this LAMP assay was 10-fold higher than regular PCR.A visual detection approach with addition of dye was feasible to interpret the positive or negative results.Conclusion: The genetic diagnostic method of Yersinia pestis based on LAMP is rapid,sensitive and specific.This method may be the potential effective way for detection of Yersinia pestis on the scene.