Legionella, IgG, IgA, IgM, BioAssay(TM) ELISA Kit

The United States Biological Legionella ELISA Test System is an enzyme-linked immunosorbent assay for the qualitative detection of total antibody (IgG/IgM/IgA) to Legionella pneumophila serogroups (1-6) in human serum. \n\nL. pneumophila was identified as the causative agent for Legionellosis (Legionella pneumonia, or Legionnaire\\'s Disease) in 1977 (1). Presently, there are more than 25 species and 33 serogroups in the family Legionellaceae, with at least 18 species associated with pneumonia, accounting for roughly 1-5% of all cases of pneumonia (2). L. pneumophila displays a multitude of morphologies including the bacillus, coccobacillus, and elongated fusiform. Although often difficult to perform, the Gram stain will be Gram-negative. The Legionella IgG/IgA/IgM ELISA test system is designed to detect antibodies to L. pneumophila in human serum. Wells of plastic microwell strips are sensitized by passive absorption with Legionella antigen. The test procedure involves three incubation steps; 1. Test serum (properly diluted) are incubated in antigen coated microwells. Any antigen specific antibody in the sample will bind to the immobilized antigen. The plate is washed to remove unbound antibody and other serum components. 2. Peroxidase Conjugated goat anti-human IgG/IgA/IgM is added to the wells and the plate is incubated. The Conjugate will react with the antibody immobilized on the solid phase in step 1. The wells are washed to remove unreacted Conjugate. 3. The microwells containing immobilized peroxidase Conjugate are incubated with peroxidase Substrate Solution. Hydrolysis of the Substrate by peroxidase produces a color change. After a period of time the reaction is stopped and the color intensity of the solution is measured photometrically. The color intensity of the solution depends upon the antibody concentration in the original test sample. \n\nThe antibody response to L. pneumophila may be both specific and nonspecific, since the patient may have antibodies to similar antigens from other Gram-negative bacteria. Optimum times for specimen collection appear to be within the first week of illness, or as soon as possible after the onset (acute specimen), and at least 3 weeks after the onset (convalescent specimen) (3). By the IFA method, a single result of >1/256 is considered presumptive evidence of legionella infection. Diagnostic titers have been reported to be absent in as many as 25% of patients (4), but the use of multiple Legionella species (5,6) as the antigen source and a polyvalent conjugate directed against IgG, IgM, and IgA (7) maximize the accuracy of serological procedures.Detection Range.\n\nDetection Range Qualitative: Positive, Negative and Cut-off\nSample: 10ul\nSpecificity: 95%\nSensitivity: 95%\nTotal Time: ~ 70 min\nShelf Life: 12-18months\n\nKit Components: \nEach kit contains the following components in sufficient quantities to perform the number of tests indicated. All reactive reagents contain 0.1% sodium azide as a preservative \n\n1. Microtiter Plate\n2. Conjugate\n3. Calibrator\n4. Positive Control\n5. Negative Control\n6. TMB\n7. Wash Buffer, 10X Diluted wash buffer to 1X\n8. Sample Diluent\n9. Stop Solution \n\nStorage and Stability:\nStore all components at 4 degrees C. Stable for 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.

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