Abstract: To detect the optimization of Vibrio vulnificus culture environment and to improve the detection rate of the bacteria. The Box-Behnken design model in Design-expert software was used to design a series of experiments with three factors at three levels in order to observe the effects of three factors such as saltness of culture medium, pH value of the nutrient solution and culture temperature on the growth velocity of Vibrio vulnificus. Based on it, a predictable mathematical model was constructed and the optimum conditions of culture medium were determined using response surface analysis (RSA) . Results showed that the optimum culture condition were:salt concentration 3.65%, pH value 6.75, culture temperature 37.00℃, resulting the theoretical value of 0.520 at OD595nm. PCR (Polymerase Chain Reaction) was applied for rapid detection of the seafood in the optimal growth environment. The obtained data indicated that vibrio vulnifius in aquatic products could be detected quickly and effectively by using the optimal culture environment and PCR.