1. Usage
The BIOKAU® Swine Foot and Mouth Disease Distinguishing ELISA Test kit, or called FMD-3ABC ELISA kit is used for detection the non-structural 3ABC-protein of swine foot and mouth disease (FMD-3ABC) antibodies in the serum, and differentiation naturally infected from vaccined swine.
2. Brief Introduction
The BIOKAU® FMD-3ABC test kit is made of the antigen coated microplate, enzyme conjugation (HRP goat-anti-pig IgG) and other reagents, which applies the indirect ELISA principle to test the antibodies against FMD-3ABC in the serum.
3. Reagents
1 3ABC Antigen coated microplate....................................... 2 pieces (96 wells)
2 Negative (green lid)/positive (red lid) serum................ 1 tube each (1 mL/tube)
3 3 ABC Enzyme conjugate (yellow lid)........................... 1 bottle (22 mL/bottle)
4 20× concentrated washing solution (white lid)............... 1 bottle (30 mL/bottle)
5 Substrate A (purple lid)/B (yellow lid).................... 1 bottle each (12 mL/bottle)
6 Stop solution (blue lid)................................................ 1 bottle (12 mL/bottle)
7 Sample diluent solution (transparent lid)....................... 1 bottle (50 mL/bottle)
4. Material Required But not Provided
Micropipettes, Distilled water, Constant temperature box or incubator, microplate-reader, Absorbent paper.
5. Warning and Procautions
1. All reagents should be adjusted to the room temperature before using the test kit.
2. Never mix up reagents from test kits in different batch number.
3. Prevent the microplate to get damp.
4. Keep dry of remaining microplates after use
6. Preparation
Dilute 20× concentrated washing solution at 1: 19 with distilled water.
7. Test Procedure
1. Take out the pre-coated microplate. Dilute the sample at 1:39 with the sample diluent and add to the wells, 100 μL each. Set 2 wells for negative control serum and 2 wells for positive control serum, 100 μL/well. Set one blank control well, add 100 μL sample diluent. Mix gently, incubate at 37℃ for 30 min.
2. Pour the liquid out of the wells, add 300 μL diluted washing solution to each well, static for 3 min, pour out. Repeat 3 times, then pat to dry on absorbent paper.
3. Add 100 μL enzyme conjugation(anti-pig IgG-HRP combination) to each well, and incubate at 37℃ for 30 min.
4. Repeat step 2.
5. Add substrate A and B one drop(50 μL) respectively to each well, mix properly, react for 10 min at room temperature at dark.
6. Add stop solution one drop (50 μL) in each well, and measure the result within 10 min.
8. Results
Set zero for the blank well, and test the OD630nm value on the microplate-reader. The conditions for the test to be tenable are that the positive control wells’ average OD630nm value is greater than or equal to 0.4, and the negative control wells’ average OD630nm value is less than 0.2.
If the sample’s OD630 value is greater than 0.4, it is judged to be positive; from 0.2 to 0.4, doubtful; and if less than 0.2, negative.
Specifications: 96 wells×2.
Expiry date: 6 months.
Storage: Storing at 4℃, in the dark.