The effects of Pseudomonas and the proteases it synthesizes in seafoods rendering them unfit for consumption are not fully recognized. A sandwich enzymelinked immunosorbent assay (ELISA) was developed wherein protease produced by Pseudomonas isolated from shrimp was used as antigen, and anti-protease IgG conjugated with alkaline phosphatase was the second antibody. Purified protease and seafood samples, naturally contaminated or artificially inoculated with Pseudomonas, were positive by ELISA. The conventional culture method took 3 days to complete, but ELISA detected Pseudomonas within 24h. The rapidity, simplicity and efficacy of this test make it useful for implementation of HACCP systems.