Intended Use:Toxoplasma IgG ELISA is intended for use in evaluating serologic status to Toxoplasma gondii infection.
Introduction:Toxoplasmosis is caused by the intracellular parasite Toxoplasma gondii and may be contracted by consuming contaminated meat or by coming in contact with cat feces containing oocysts. In adolescence and adulthood, most infections are subclinical. However, if a pregnant woman contracts toxoplasmosis, it may be passed through the placenta to the fetus, resulting in congenital toxoplasmosis, which is a cause of mortality and malformation. Asymptomatic infants may develop abnormalities later in life.
Test Principle:Purified Toxoplasma gondii antigen is coated on the surface of microwells. Diluted serum is added to the wells, and the Toxoplasma gondii IgG-specific antibody, if present, binds to the antigen. All unbound materials are washed away. HRP-conjugate is added, which binds to the antibody-antigen complex. Excess HRP-conjugate is washed off and a solution of TMB Reagent is added. The enzyme conjugate catalytic reaction is stopped at a specific time. The intensity of the color generated is proportional to the amount of IgG-specific antibody in the sample. The results are read by a microwell reader compared in a parallel manner with calibrator and controls.
Kit Components:T8075-51T1: Antigen-Coated Wells (1 plate, 96 wells, 12x8 strips), Microtiter Wells coated with purified Toxo antigenT8075-51T2: Enzyme Conjugate Reagent (12ml). Contains goat anti-human IgG conjugated to horseradish peroxidase with preservativesT8075-51T3: Sample Diluent (22ml). Contains phosphate buffer with preservativesT8075-51T4: Calibrator 1: 0 IU/ml (100ul/vial), Contains Toxo IgG negative serum with preservatives. Natural cap
T8075-51T5: Calibrator 2: 32 IU/ml (100ul/vial), Contains Toxo IgG positive serum with preservatives. Yellow cap
T8075-51T6: Calibrator 3: 100 IU/ml (100ul/vial), Contains Toxo IgG positive serum with preservatives. Red cap
T8075-51T7: Calibrator 4: 300 IU/ml (100ul/vial), Contains Toxo IgG positive serum with preservatives. Green cap
T8075-51T8: Low Control (100ul/via). Range stated on label. Blue cap
T8075-51T9: High Control (100ul/vial). Range stated on label. Purple cap
Storage and Stability:Store all components at 4°C. Stable for at least 6 months. For maximum recovery of product, centrifuge the original vial prior to removing the cap.
Assay Procedure:1. Place the desired number of coated wells into the holder. 2. Prepare 1:40 dilution of test samples, calibrators, and controls by adding 5ul of the sample to 200ul of Sample Diluent. Mix well. 3. Dispense 100ul of diluted sera, calibrators, and controls into the appropriate wells. For the reagent blank, dispense 100ul Sample Diluent in 1A well position. Tap the holder to remove air bubbles from the liquid and mix well. 4. Incubate at 37C for 30 minutes. 5. At the end of incubation period, remove liquid from all wells. Rinse and flick the microtiter wells 5 times with diluted Wash Buffer (1x). 6. Dispense 100ul of Enzyme Conjugate to each well. Mix gently for 10 seconds. 7. Incubate at 37C for 30 minutes
8. Remove Enzyme Conjugate from all wells. Rinse and flick the microtiter wells 5 times with diluted Wash Buffer (1x). 9. Dispense 100ul of TMB Reagent into each well. Mix gently for 10 seconds. 10. Incubate at 37C for 15 minutes. 11. Add 100ul of Stop Solution (1N HCl) to stop reaction. 12. Mix gently for 30 seconds. It is important to make sure that all the blue color changes to yellow color completely. Note: Make sure there are no air bubbles in each well before reading. 13. Read O.D. at 450 nm within 15 minutes with a microwell reader.
