Serological diagnosis of infectious diseases that generate a highly heterogeneous antibody repertoire, such as tuberculosis, requires tests based on cocktails of antigens. We describe a new method called multi-antigen print immunoassay (MAPIA) for cocktail-based serological diagnosis. The assay entails the application of antigen to nitrocellulose membranes by micro-aerosolization (printing), followed by antibody detection using standard chromogenic immunodevelopment. Cocktails of protein antigens of Mycobacterium tuberculosis tested by MAPIA were found to maintain the serological activity of each of their components. In contrast, the same cocktails tested by enzyme-linked immunosorbent assay (ELISA) had a serological activity that was lower than the sum of the activities of their components. Consequently, cocktail-based MAPIA attained the diagnostic sensitivity expected on the basis of single antigen results, while a significant loss of diagnostic sensitivity was observed with cocktail-based ELISA. Thus, the MAPIA format is superior to conventional ELISA for the serological diagnosis of infectious diseases characterized by heterogeneous antibody responses.