Method Description

The ZIKV Detect 2.0 IgM Capture enzyme-linked immunosorbent assay (ELISA) is for the detection of human IgM antibodies targeting the Zika virus (ZIKV) envelope glycoproteins. Polystyrene microtiter wells are precoated with polyclonal capture antibodies against human IgM. Positive control, negative control, and patient serum samples are diluted into a sample dilution buffer and then added to the ELISA plate in appropriate locations. After incubation and washing, a subsequent ready-to-use (RTU) ZIKV antigen (Zika Ag), a cross-reactive control antigen, and a normal cell antigen are added separately to each corresponding well. After incubation and washing, a RTU secondary antibody solution is added to each well. After a subsequent incubation and wash steps, an enzyme conjugate solution comprising horseradish peroxidase-labeled antimouse antibody is added to each well. After washing, wells are incubated with a tetramethylbenzidine substrate. An acidic stop solution is then added, and the degree of enzymatic turnover is determined by the absorbance (optical density) measurement at 450 nanometers. If human IgM antibodies targeting the ZIKV envelope glycoproteins are present, a complex is formed consisting of the IgM, antigen, secondary antibody, and conjugate. If IgM antibodies targeting the ZIKV envelope glycoproteins are not present, then the antigen, antibody, and conjugate are washed away.(Package insert: InBios ZIKV Detect 2.0 IgM Capture ELISA. InBios International, Inc; 09/23/2021)