Herpes Simplex Virus 2 IgM ELISA

The Herpes Simplex Virus Type 2 IgM ELISA Kit is a solid phase enzyme-linkedimmunosorbent assay (ELISA).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 fromspecific IgG and to remove rheumatoid factors. This pretreatment avoids false negative or falsepositive results. Microtiter wells as a solid phase are coated with HSV-2 antigen.Diluted specimens and ready-for-use controls are pipetted into these wells. During incubationHSV-2-specific antibodies of positive specimens and controls are bound to the immobilizedantigens.After a washing step to remove unbound sample and control material horseradish peroxidaseconjugated antihuman IgM antibodies are dispensed into the wells. During a second incubationthis anti-IgM conjugate binds specifically to IgM antibodies resulting in the formation of enzymelinked immune complexes.After a second washing step to remove unbound conjugate the immune complexes formed (incase of positive results) are detected by incubation with TMB substrate and development of ablue color. The blue color turns into yellow by stopping the enzymatic indicator reaction withsulfuric acid.The intensity of this color is directly proportional to the amount of HSV-2-specific IgM antibody inthe specimen. Absorbance at 450 nm is read using an ELISA microtiter plate reader.