To develop a more economical, simple and rapid detecting and differentiating Vibrio cincinnatiensis Differential Medium (VciDM), V. cincinnatiensis is one species of the major pathogens in seafood. Based on the specific enzyme systems and the corresponding metabolisms of V. cincinnatiensis, according to the resistibility to different antibacterial ingredients, VciDM was designed for the rapid separation, identification or detection of V. cincinnatiensis. The tests of color effects, plating efficiency, sensitivity and specificity were done to evaluate the efficiency of VciDM. The results showed that: VciDM had good selectivity and specificity. After incubation for 16?24 h at 37°C ± 0.1°C, the V. cincinnatiensis formed yellow colonies on VciDM. The other species of bacteria did not form colonies or formed green colonies on VciDM. VciDM showed a mean plating efficiency of V. cincinnatiensis cells of (82.14 ± 4.45)% and its detection limit was 101 CFU/mL. As compared with the conventional methods, VciDM used for isolating and detecting V. cincinnatiensis, which is simpler operation, lower cost, more convenient for observation, higher sensitivity and specificity. Accordingly, it is suitable for kinds of inspection agencies to detect V. cincinnatiensis, especially for the use in rapid detecting and monitoring V. cincinnatiensis in seafood in the primary production and processing plants. VciDM can be used to rapidly and initiatively detect V. cincinnatiensis, which is usefull to rake quick infection control measurees.