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This invention discloses primers, kits, methods, and applications for detecting pathogenic *Pseudomonas proteus* strains based on the *oprL* gene, belonging to the field of microbial detection technology. This invention provides specific primers for detecting pathogenic *P. proteus* strains based on the *oprL* gene. The primers are designed based on the differential patterns identified after multiple sequence alignment, specifically by analyzing the results of multiple sequence alignments of all *P. proteus* *oprL* gene sequences in the NCBI database. The nucleotide sequences of the primers are shown in SEQ ID No. 1, SEQ ID No. 2, and SEQ ID No. 3. The *oprL* gene of *P. proteus* is amplified by PCR, and the PCR amplification products are then subjected to electrophoresis. The pathogenicity of *P. proteus* can be effectively distinguished based on the electrophoresis image, enabling rapid detection of *P. proteus* infecting large yellow croaker. The primers have good specificity, and the detection method is simple and intuitive.