Study on loop-mediated isothermal amplification (LAMP) technique for detection of <I>Vibrio parahaemolyticus</I> based on the<I>gyrB</I> gene

ObjectiveTo establish a method of loop-mediated isothermal amplification to detect IVibrio parahaemolyticus/I for the laboratory at grassroots unit. MethodsFour primers (two inner primers, two outer primers) were designed specifically to recognize the IgyrB/I gene of IVibrio parahaemolyticus/I. ResultsIt only takes 1.5 h to conduct the detection with optimized condition and the result can be judged by naked eyes or electrophoresis. Three strains of different species IV. Parahaemolyticus/I, 23 local strains of IV. parahaemolyticus/I and 32 strains of other intestinal bacteria were detected by the method with high specificity. The method was sensitive with the lowest detectable limit (LDL) of 24　cfu/ml. In the field detection of 60 shellfishes, the positive rate was 100%, which was higher than that of traditional method and was consistent with the result of RT-PCR. ConclusionThe method is suitable for the laboratory at grassroots unit, emergency detection and field detection, which is rapid, sensitive, specific, simple, economical, which and is worth to spread.