Abstract:[Objective] The visual antibody array for rapidly detecting Escherichia coli O157:H7 and Samonella typhimurium was built. [Methods] Combined with immunology and protein chip technology, Based on double antibody sandwich assay principle. Used high-throughput protein chip and visualization technology, the method for simultaneous detecting E. coli O157:H7 and Salmonella typhimurium has been selected, and only one sample was used. [Results] The test results couble be visible to the naked eye, the detection of time was as short as 90 min. The sensitivity of pure broth was 105 CFU/mL and artificially contaminated food was 106 CFU/mL. The sensitivity was coincided with conventional ELISA and had good detection specificity and repeatability. [Conclusion] The visual antibody array couble be examined by naked eye, which owned high-throughput detection, easy operation, low cost of detection and didn’t need large equipment. The study showed a satisfactory prospect and provided a new rapid assay for detection of Pathogenic microorganisms.