Lyme Disease, Human, IgM, BioAssay(TM) ELISA Kit

Principle of the Assay:\nThis Borrelia burgdorferi ELISA test system is designed to detect IgM class antibodies to B. burgdorferi in sample serum. Wells of plastic microwell strips are sensitized by passive absorption with B. burgdorferi antigen. The test procedure involves three incubation steps.\n1. Test serum are diluted with the Sample Diluent provided. The Sample Diluent contains anti- sample IgG which precipitates and removes IgG and rheumatoid factor from the sample leaving IgM free to react with the immobilized antigen. During sample incubation, any antigen specific IgM antibody in the sample will bind to the immobilized antigen. The plate is washed to remove unbound antibody and other serum components. \n2. Peroxidase Conjugated goat anti-sample IgM (mu chain specific) is added to the wells and the plate is incubated. The Conjugate will react with IgM antibody immobilized on the solid phase in step 1. The wells are washed to remove unbound Conjugate. \n3. 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\nKit Components:\nAll reactive reagents contain sodium azide as a preservative at a concentration of 0.1%. \nL8001-21A: Microtiter Plate: 1x96wells, coated with capture antibody. \nL8001-21B: IgM (mu) (HRP) Pab Gt x Hu, mu chain specific, 1x15ml\nL8001-21C: Positive Control, (Human Serum), 1x350ul\nL8001-21D: Calibrator (Human Serum), CF=0.43, 1x500ul\nL8001-21E: Negative Control (Human Serum), 1x350ul\nL8001-21F: IgG Pab Gt x Hu , 1x12ml\nL8001-21G: Sample Diluent, 1x30ml, (Contains PBS, pH7.2, BSA, Tween-20)\nL8001-21H: Wash Buffer, 10X, 1x100ml (Contains PBS, Tween-20) \nL8001-21J: Tetramethylbenzidine (TMB): 1x15ml (Contains <15% DMSO) \nL8001-21K: Stop Solution:1x15ml (1M H2SO4, 0.7M HCl) \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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SPECIFICATIONS

Catalog Number

L8001-21

Size

1Kit

Applications

ELISA

References

1. Steere, A.C., et al., J. Infect. Dis. 154: 295-300 (1986). 2. Rosenfeld, M.E.A.:Serodiagnosis of Lyme disease. J. Clin. Microbiol. 31: 3090-3095 (1993). 3. Steere, A.C., et al., New Engl. J. Med. 308: 733 (1983). 4. Bakken, L.L., et al., J. Clin. Microbiol. 35: 537-543 (1997). 5. U.S. Department of Labor (OSHA):Occupational Exposure to Bloodborne Pathogens. Final Rule. 21CFR 1910.1030. 6. Procedures for the Handling and Processing of Blood Specimens: NCCLS Procedure H18. Approved guideline. 7. Procedures for the collection of diagnostic blood specimens by venipuncture:NCCLS Procedure H3, Approved Standard. 8. Barbour A: Laboratory Aspects of Lyme Borreliosis. Clin Micr. Rev. 1: 399-414 (1988). 9. Russel, H., et al., J. Infect. Dis. 149: 465 (1984). 10. Magnarelli, L.A., et al., J. Infect. Dis. 156: 183-188 (1987). 11. Hunter, E.F., et al., Sex. Trans. Dis. 13: 236 (1986). 12. Shrestha, M., et al., Am. J. Med. 78: 235 (1985). 13. Steere, A.C., et al., Ann. Intern. Med. 99: 22 (1983). 14. Craft, J.E., et al., Yale J. Biol. Med. 57: 561 (1984). 15. Reik ,L., et al., Neurology 35:267 (1985).