Loop-mediated isothermal amplification (LAMP) was applied to establish a rapid duplex LAMP-dissociation curve method for the simultaneous detection of toxigenic Vibrio cholerae and Vibrio parahaemolyticus in foods. The primers were designed according to the sequences of the ctxA gene of Vibrio cholerae and the gyrB gene of Vibrio parahaemolyticus, and the amplification products were analyzed by dissociation curves to determine the target bacteria using the DNA template. The results from the detection of nine target bacterial strains and 17 non-target bacterial strains were consistent with the expected results, and the target bacteria represented by the DNA template could be accurately analyzed using the characteristic peaks in the dissociation curve. DNA sequencing results of LAMP products indicated that the amplified sequences were also consistent with the target gene sequences, further confirming the specificity of this LAMP method. The experimental results showed that the detection limits for the two target bacteria could reach 100 fg DNA/tube. The results suggest that the established method has a high specificity, and can be used as an important technique for the rapid detection of two pathogenic Vibrio species in foods.