Rapid Detection of Vibrio parahaemolyticus by Loop-mediated Isothermal Amplification Method

A pair of outer primers and a pair of inner primers were specially designed for recognizing six distinct sequences on target tlh gene by loop-mediated isothermal amplification of DNA (LAMP) method. The sensitivities of LAMP method and PCR were conpared for specific detection of Vibrio parahaemolyticus. The results showed that target DNA was amplified and visualized ladder-like pattern of bands on agarose gel by the detection system within 60 min at isothermal temperature 65 ℃; White precipitate allows easy and rapid detection of target DNA by naked eyes; The resulting amplicons are visualized by adding SYBR Green I to the reaction tube; The LAMP assay has a detection limit of 10 CFU/ml, while that of PCR is 103 CFU/ml, that is to say that the sensitivity of LAMP is 100 times of that of PCR. So the LAMP method reported here demonstrates a potential and valuable means for detection of Vibrio parahaemolyticus especially for its rapidity, simplicity and low cost. The only equipment needed for the amplification reaction is a regular laboratory water bath or heat block that furnishes a constant temperature. The LAMP assay is particularly suitable for rapid clinical diagnosis.