Epstein-Barr Virus (EBV) – [VCA] IgM

The Epstein-Barr Virus (VCA) IgM ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA) Specimen samples are diluted with Sample Diluent and additionally incubated with IgG-RF-Sorbent, containing hyper-immune anti-human IgG-class antibody to eliminate competitive inhibition from specific IgG and to remove rheumatoid factors. This pretreatment avoids errors in results. Microtiter wells as a solid phase are coated with inactivated Epstein-Barr viral capsid antigen. Pretreated sample specimens and ready-for-use controls are pipetted into these wells. During incubation Epstein-Barr viral capsid antigen-specific antibodies of specimens where analyte is present and controls are bound to the immobilized antigens. After a washing step to remove unbound sample and control material horseradish peroxidase conjugated antihuman IgM antibodies are dispensed into the wells. During a second incubation this anti-IgM conjugate binds specifically to IgM antibodies resulting in the formation of enzyme-linked immune complexes. After a second washing step to remove unbound conjugate the immune complexes formed (in cases where analyte is found) are detected by incubation with TMB substrate and development of a blue color. The blue color turns into yellow by stopping the enzymatic indicator reaction with sulfuric acid. The intensity of this color is directly proportional to the amount of Epstein-Barr viral capsid antigen-specific IgM antibody in the sample specimen. Absorbance at 450 nm is read using an ELISA microtiter plate reader.