Borrelia burgdorferi IgG ELISA
The IMMUNOLAB Borrelia burgdorferi IgG antibody ELISA kit has been designed for the detection and the quantitative determination of specific IgG antibodies against Borrelia burgdorferi in serum and plasma. Further applications in other body fluids are possible and can be requested from the Technical Service of IMMUNOLAB.
Borrelia burgdorferi belongs to the family of spirochetes, of which three types have been identified to be human pathogenic: Borrelia burgdorferi sensu stricto, B. afzelii and B. garinii. The endemic areas of Borrelia are in Central and Eastern Europe, as well as Russia, China and Japan. The illness is transferred via tick bites, in Europe mainly by Ixodes ricinus. In the endemic zones, like Southern Germany and Austria, up to 50 % of the ticks are infected.
In the clinical course, after an erythema migrans, e.g. with neuroborreliosis, which appears at the first stage, also chronical arthritis, encephalitis, meningitis, myositis and hepatitis are observed. Treatment is done via different antibiotics, e.g. doxycyclin, amoxicillin, cefuroxim and penicillin G. A specific immunization is possible with immunoprophylaxis either by a recombinant OspA or by a recombinant polyvalent OspC vaccine.
The laboratory diagnosis is performed by the detection of antibodies in blood and cerebrospinal fluid. Methods employed are: ELISA, immunofluorescence, hemagglutination or Western blot. Besides whole cell extracts, recently there are increasingly used purified or recombinant single proteins as antigens. This brings however generally a decrease in sensitivity. It could be shown that between the various test methods there appear significant differences in the interpretation, so that the most reliable method seems to be the follow-up of the titer development. Western blot serves as a confirmatory test, because electrophoretically separated single antigens can be evaluated in their reaction with specific serum antibodies.
The IMMUNOLAB Borrelia burgdorferi IgG ELISA test kit contains besides a whole cell antigen extract of Borrelia burgdorferi sensu stricto, which cross-reacts with Borrelia afzelii and Borrelia garinii, an addition of pure OspC, which increases the specificity and sensitivity of the assay. If the test shows only a positive IgG result (IgM negative), the immunity of the patient can be concluded. As mentioned above, in the case of a clearly positive clinical finding, there should be performed a confirmation of the titer increase with the same assay. With a positive IgM (IMMUNOLAB ELISA ILE-BOR03) at the same time, there is a strong suspicion of an active disease.
The IMMUNOLAB Borrelia burgdorferi IgG antibody test kit is based on the principle of the enzyme immunoassay (EIA). Borrelia burgdorferi antigen is bound on the surface of the microtiter strips. Diluted patient serum or ready-to-use standards are pipetted into the wells of the microtiter plate. A binding between the IgG antibodies of the serum and the immobilized Borrelia burgdorferi antigen takes place. After a one hour incubation at room temperature, the plate is rinsed with diluted wash solution, in order to remove unbound material. Then ready-to-use anti-human-IgG peroxidase conjugate is added and incubated for 30 minutes. After a further washing step, the substrate (TMB) solution is pipetted and incubated for 20 minutes, inducing the development of a blue dye in the wells. The color development is terminated by the addition of a stop solution, which changes the color from blue to yellow. The resulting dye is measured spectrophotometrically at the wavelength of 450 nm. The concentration of IgG antibodies is directly proportional to the intensity of the color.
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