Intended Use:
The Syphilis TPA-Treponema pallidum IgM ELISA kit provides materials for the qualitative and semi quantitative determination of IgM-class antibodies to Treponema pallidum in serum. This is intended for in vitro diagnostic use only
Principle of the Assay:
The Treponema pallidum IgM ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA). Sample Diluent is used to dilute patient samples, and then incubating with IgG-RF-Sorbent, containing hyper-immune anti-human IgG-class antibody to exclude competitive inhibition from specific IgG, and to remove rheumatoid factors. This pre-treatment avoids false negative or false positive results. Microtiter wells as a solid phase are coated with Treponema pallidum antigen. Pre- treated patient specimens and ready-to-use controls are pipetted into these wells. During incubation, Treponema pallidum-specific antibodies of positive specimens and controls are bound to the immobilized antigens.
After the washing step, that removes unbound sample and controls material HRP- conjugated anti-human IgM antibodies are then dispensed into the wells. During the second incubation, this anti-IgM conjugate binds specifically to IgM antibodies that forms enzyme-linked immune complexes. After a second washing step to remove unbound conjugate, the immune complexes formed (in case of positive results) are detected by incubation with TMB substrate and a blue color develops. The blue color turns to yellow by stopping the enzymatic indicator reaction with sulfuric acid. The intensity of this color is directly proportional to the amount of Treponema pallidum- specific IgM antibody in the patient
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