Borrelia burgdorferi belongs to the family of spirochetes, of which three types have been identified to be pathogenic to humans: Borrelia burgdorferi sensu stricto, B. afzelii and B. garinii. The endemic areasof 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 endemic zones, like Southern Germany and Austria, up to 50% of ticks are infected. In the clinical course, after an erythema migrans, e.g. with neuroborreliosis, which appears at the first stage, chronical arthritis, encephalitis, meningitis, myositis and hepatitis are also observed. Treatment is usually by different antibiotics, e.g. doxycyclin, amoxicillin, cefuroxim and penicillin G. Specific immunization is possible with immunoprophylaxis either by a recombinant OspA or by a recombinantpolyvalent OspC vaccine. 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, purified or recombinant single proteins are increasingly used as antigens. This generally results however in a decrease in sensitivity. It has been shown that there are significant differences between the various test methods used for interpretation and the most reliable method seems to be the follow-up of titer development. Western blot serves as a confirmatory test, because electrophoretically separated single antigens can be evaluated in their reaction with specific serum antibodies.
Intended Use:The Borrelia burgdorferi IgM antibody ELISA kit has been designed for the detection and the quantitative determination of specific IgM antibodies against Borrelia burgdorferi in serum and plasma.
Sample Type:Principally serum or plasma (EDTA, heparin) can be used for the determination. Lipemic, hemolytic or bacterially contaminated samples can cause false positive or false negative results. For the performance of the test the samples (not the standards) have to be diluted 1:101 with readyto-use sample diluent (e.g. 5ul serum + 500ul sample diluent).
Sensitivity:0.99U/ml
Test Principle:The Borrelia burgdorferi IgM 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 serum or ready-to-use standards are pipetted into the wells of the microtiter plate. A binding between the IgM antibodies of the serum and the immobilized Borrelia burgdorferi antigen takes place. After 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-IgM 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 a wavelength of 450nm. The concentration of IgM antibodies isdirectly proportional to the intensity of the color. The Borrelia burgdorferi IgM ELISA test kit contains both a whole cell antigen extract of Borrelia burgdorferi sensu stricto (which cross-reacts with Borrelia afzelii and Borrelia garinii) and pure OspC, which greatly increases the specificity and sensitivity of the assay.
Crossreactivity:No cross-reactivity to TBEV (FSME)
Precision:Intra-Assay: 5%Inter-Assay: 9%
Recovery:110-111%
Linearity:77-120%
Interferences:No interferences to bilirubin up to 0.3mg/ml, hemoglobin up to 8mg/ml and triglycerides up to 5mg/ml.
Kit Components:Borrelia burgdorferi antigen coated microtiter strips, 1x12Calibrator A (Negative Control), 1x2mlCalibrator B (Cut-Off Standard), 1x2mlCalibrator C (Weak Positive Control), 1x2mlCalibrator D (Positive Control), 1x2mlEnzyme Conjugate, 1x15mlSubstrate, 1x15mlStop Solution, 1x15mlSample Diluent, 1x60mlWashing Buffer (10x), 1x60ml
Storage and Stability:Store powder at 4°C liquid at -20°C. Store other components at 4°C. Stable for at least 6 months For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
