Brucella IgM ELISA, 96 wells

Introduction
The DRG Brucella IgM Enzyme Immunoassay Kit provides materials for the qualitative and semiquantitative
determination of IgM-class antibodies to Brucella in serum. This assay is intended for in vitro use only.

Principle Of The Test
The DRG Brucella IgM ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA)
Patient samples are diluted with Sample Diluent and additionally incubated with IgG-RF-Sorbent, containing hyperimmune
anti-human IgG-class antibody to eliminate competitive inhibition from specific IgG and to remove rheumatoid
factors. This pretreatment avoids false negative or false positive results.
Microtiter wells as a solid phase are coated with Brucella antigen.
Diluted patient specimens and ready-for-use controls are pipetted into these wells. During incubation Brucella-specific
antibodies of positive specimens and controls are bound to the immobilized antigens.
After a washing step to remove unbound sample and control material horseradish peroxidase conjugated anti-human 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 case of positive results) 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 Brucella-specific IgM antibody in the patient
specimen. Absorbance at 450 nm is read using an ELISA microtiter plate reader.