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For Farm Ranch Breeding Grounds Cattle Bovine Tuberculosis Animal Rapid Test Kit
|Cattle&Sheep Test Kit
|Taeing+Pit Microchip Reader+Wholesale Manufacturer
|US $ 0.9 / piece
|Terms of Payment
The Bovine Tuberculosis (BTB) Antibody Test Kit is designed to detect antibodies to Bovine Tuberculosis in bovine serum and is used as a serological aid in the evaluation of the detection of tuberculosis caused by Mycobacterium bovis.
The kit uses an indirect ELISA method in which enzyme-labeled lath microtiter wells are pre-encapsulated with bovine Mycobacterium tuberculosis antigen. In the test, diluted serum to be tested is added, and after warming, if the sample contains antibodies specific for Mycobacterium tuberculosis, it binds to the antigen on the coated plate, and after washing to remove unbound antibodies and other components, the enzyme secondary antibody is added to specifically bind to the antigen-antibody complex on the detection plate, and then the unbound enzyme conjugate is removed by washing, and the TMB substrate solution is added to the microtiter wells, and the enzyme After the reaction was terminated by adding the termination solution, the absorbance A value in the reaction wells was measured at 450 nm with an enzyme marker.
Equipment and reagents to be provided
1. Micro pipettes: 0.5µl-10µl, 10µl-100µl, 100µl-1000µl. 2.
2. Disposable pipette tips. 3.
3. Volumetric cylinder: 500ml.
4. 96-well plate enzyme marker.
5. distilled water or deionized water. 6.
6. Wash bottle or plate washer.
Take the whole blood of the animal and prepare the serum according to the conventional method, requiring the serum to be clear and free of hemolysis.
Preparation of washing solution
Concentrated washing solution should be restored to room temperature before use, and if salt crystals, shake to dissolve the crystallized salt, and then dilute 10 times with distilled or deionized water. The diluted washing solution can be stored at 4℃ for about a week.
Dilution of samples
Dilute the serum to be tested in serum dilution plates at a volume ratio of 1:99 (e.g., 495 μl of sample dilution with 5 μl of serum to be tested).
Note: Negative and positive controls do not need to be diluted. Change the tips after taking each sample and record accurately the position of each sample on the plate. Each sample should be well mixed before adding to the microtiter wells of the coated plate.
1. Each reagent in the kit should be equilibrated to room temperature before use, shaken well before use, and put back to 2-8°C after use.
2. The reagent components of different varieties and batches of kits should not be mixed, and reagent contamination should be prevented when using reagents.
3. The substrate and termination solution may be irritating to the skin and eyes, and care should be taken to protect them when using them.
4. TMB (color developing solution) should not be exposed to strong light and avoid contact with oxidizing agents.
5. Avoid moisture or water after unpacking the assay plates (unused antigen-coated plates with desiccant should be placed in self-sealing bags and placed at 2-8°C as soon as possible).
6. All waste should be disposed of reasonably before discarding to avoid contamination.
7. The best results can be obtained by strictly following the operating instructions. The entire process of pipetting, timing and washing during operation must be precise.
8. Serum dilution plates are disposable and should not be reused; the maximum capacity of serum dilution plates is 300 μl/well.
1. Take the pre-wrapped assay plate (according to the number of samples, it can be disassembled and used separately) and add 100μl of the diluted serum to the wells of the assay plate. At the same time, set up 1 negative control well and 2 positive control wells, and add 100 μl of each negative and positive control into the wells to gently shake the sample in the wells (do not overflow).
2. Cover with sealing film (can be cut according to actual needs) and incubate at 37℃ for 30 minutes.
3. Carefully remove the sealing film and shake off the solution from the wells. Add 250 ul of working concentration washing solution to each well, then shake off the liquid in the wells.
4. Add 100μl of enzyme conjugate to each well, cover with sealing membrane and incubate at 37℃ for 30 minutes.
5. Carefully remove the sealing membrane, shake off the solution from the wells and wash 4-6 times as in 3. Remember to pat dry on clean blotting paper at the end.
6. Add 100 µl of color development solution to each well, shake lightly and mix well, cover the sealing membrane and develop the color for 10 min at 37°C with closed light.
7. Add 50 µl of termination solution to each well to terminate the reaction and determine the results within 10 minutes.
Determination of results
The absorbance OD value was read at 450nm (630nm as reference wavelength) using an enzyme marker.
The test was set up under the following conditions
The OD value of negative control (N) is <0.2, while the OD value of positive control (P) is >0.4.
sample OD value ÷ positive control OD mean value = S/P value
Negative and positive judgments:
S/P<0.3 was judged as negative.
S/P≥0.3 was judged as positive.
Storage method: 2 - 8ºC.
Expiration date: 12 months.