Abstract: Plesiomonas shigelloides strain LCCiL90625NA, isolated from diseased Ctenopharyngodon idellus, was used for rabbit-serum preparation by injecting the formalin-inactivated bacteria. The rabbit immunoglobulin G was purified by Protein A column. An indirect ELISA was developed for detection of LCCiL90625NA, using rabbit polyclonal antibodies against P. shigelloides as detecting antibody, the titers of that could reach to 1﹕1.28×105. Indirect ELISA assay indicated that the polyclonal antibody had a specific and sensitivity detection for P. shigelloides. The ELISA had no cross-reaction with Vibrio parahaemolyticus, V. vulnificus, V. alginolyticus, V. cholera non O-1, Aeromonas hydrophila, A. caviae, A. sobria, A. trota and A. salmonicida，and the minimum amount of P. shigelloides could be detected by this ELISA, was 1.0×104 CFU?mL-1. The results revealed that the ELISA method was quick，sensitive and repeatable，which was applicable to the detection of P. shigelloides in Ctenopharyngodon idellus．