The invention discloses an on-site detection immuno-chip, comprising a chip carrier and an agarose gel layer which is paved on the chip carrier, wherein, the agarose gel layer is fixed with a plurality of antibody microarrays of 4*4, and chip-dedicated fences or Super PAP Pen scribings are used for separating the microarrays from each other; the double antibody sandwich principle is adopted to detect the antigen to carry out the dot matrix of corresponding capture antibody, positive control and negative control on the solid phase carrier simultaneously; the antibody protein is connected with the solid phase carrier through the covalent bond and physical adsorption; the sample liquid to be detected and the chip are directly incubated; the antigen to be detected in the samples combine with the corresponding antibody which is fixed on the chip; a specific monoclonal antibody probe marked by horse radish peroxidase is added and the macroscopic detection results can be obtained after the coloration of substrates. The invention can detect viruses in multiple aquatic animal samples and the results are macroscopic, thus being applicable to rapid and accurate detection of viruses of aquatic animals in breeding production.