Immunoassay platform for the serological discrimination of closely related viruses

An immunoassay for discriminating non-human antibodies specific for a target virus from non-human cross-reacting antibodies to related viruses in a single sample, wherein the target virus comprises a flavivirus, a coronavirus, a herpes virus, an influenza virus, or a hepatitis virus, the immunoassay comprising:

• (a) immobilizing target virus antigens on each of a first immunoassay substrate and a second immunoassay substrate,
• (b) introducing the non-human sample comprising antibodies to each of the first immunoassay substrate and the second immunoassay substrate on which the target virus antigens have been immobilized;
• (c) (i) introducing blocking antibodies of origins other than the non-human species against target virus antigens to the first immunoassay substrate and introducing a blank solution to the second immunoassay substrate,
• or alternatively,
• (ii) providing the first immunoassay substrate with blocking antibodies preloaded thereon, with or without introducing a blank solution to the second immunoassay substrate,
• so that the blocking antibodies compete with the non-human species antibodies in the non-human species sample to bind with the immobilized target virus antigens on the first immunoassay substrate, and the non-human species antibodies of the non-human species sample bind with the immobilized target virus antigens on the second immunoassay substrate in the absence of the blocking antibodies;
• (d) introducing a non-human species antibody detection reagent to each of the first and second immunoassay substrates, wherein the non-human species antibody detection reagent binds to non-human species antibody that remain on the first immune assay substrate and the second immunoassay substrate;
• and
• (e) generating a first output signal correlative of a first amount of non-human antibodies bound with target virus antigens immobilized on the first immunoassay and generating a second output signal correlative of a second amount of non-human antibodies bound with target virus antigens immobilized on the second immunoassay,
• wherein a predetermined higher difference level between the first and second output signals contraindicates the presence of non-human antibodies to target virus, and a predetermined lower difference level between the first and second output signals indicates the presence of non-human antibodies to the target virus.