Rickettsia conorii IgM ELISA

The ELISA test microwells in this kit utilize the immunodominant outer membrane protein (rOmpB), which contain both species-specific and more broadly reactive determinants. Antigens used in this assay were purified from Rickettsia conorii, yet react much like antigens from Rickettsia rickettsii, Rickettsia slovaka and Rickettsia africae.

Specimen sample sera are diluted in a Sample Diluent and incubated in the coated microwells to allow binding of serum antibody to the solid-phase antigens. The microwells are then washed to remove unreacted serum proteins, and an enzyme-labelled anti-human IgG (Enzyme Conjugate) is added to label the bound antibody. After an incubation period, the microwells are washed to remove unbound Enzyme Conjugate. An enzyme substrate (TMB Substrate) is then added to quantitate the bound peroxidase portion of the Conjugate.

Development of a blue color is directly proportional to the amount of reactive serum antibody. This timed reaction is interrupted with a Stop Solution that turns the blue reactions to yellow and stabilizes the final color intensity. Color intensity (Absorbance) is measured at a wavelength of 450 nm on a microtiter plate reader or spectrophotometer.