The JE (Japanese Encephalitis) IgM Capture ELISA test for exposure to Japanese Encephalitis Virus (JEV) is an ELISA assay system for the detection of IgM antibodies in human serum to JEV-derived recombinant antigen (JERA) (1-4). The JE (Japanese Encephalitis) IgM Capture ELISA consists of one enzymatically amplified two-step sandwich-type immunoassay. In this assay, JE Negative Control (represents non-reactive serum), JE IgM Positive Control (represents reactive serum), and unknown serum samples are diluted with Sample Dilution Buffer, then incubated in microtitration wells which have been coated with anti-human IgM antibodies. This is followed by incubation with both JEV-derived recombinant antigen (JERA) and Normal Cell Antigen (NCA) separately. After incubation and washing, the wells are treated with a JERA-specific antibody labeled with the enzyme horseradish peroxidase (HRP). After a third incubation and washing step, the wells are incubated with the tetramethylbenzidine (TMB) substrate. An acidic stopping solution is then added and the degree of enzymatic turnover of the substrate is determined by absorbance measurement at 450 nanometers. Above a certain threshold, the ratio of the absorbencies of the JERA and the control wells accurately determines whether antibodies to JEV are present.