Packaging specifications

96T × 1/box, 96T × 2/box, 96T × 5/box

Principle and Applications

This kit consists of an enzyme-linked immunosorbent assay (ELISA) plate pre-coated with Brucella antigen, enzyme labels, and other accompanying reagents. It employs the ELISA principle to detect Brucella antibodies in animal serum and plasma samples. It can be used for evaluating the efficacy of Brucella vaccines and for auxiliary diagnosis.

Composition of the kit

Sample Preparation

1. Collect whole blood from animals and prepare serum using conventional methods. The serum must be clear, free of hemolysis, and free of contamination. Samples can be stored at 2–8℃ for up to one week; for long-term storage, they should be kept at -20℃.

2. First, dilute the serum to be tested 20-fold using the sample diluent (e.g., add 190 μL of sample diluent to a dilution plate, then add 10 μL of serum and mix thoroughly). The negative and positive controls do not need to be diluted.

3. Before use, the concentrated detergent should be brought to room temperature to dissolve any precipitates. Then, dilute it 20-fold with distilled water or deionized water to prepare the working detergent solution (e.g., 19 parts distilled water or deionized water + 1 part concentrated detergent).

Inspection method

1. Before use, place the kit at room temperature for 30 minutes to allow it to return to room temperature.

2. Take the required number of ELISA plate strips, set up 1 well for the blank control and 2 wells each for the negative and positive controls. Seal any unused strips promptly and store them at 2–8°C.

3. Add 100 μL of sample diluent to the blank control well; add 100 μL of negative and positive controls respectively to the negative and positive control wells; and add 100 μL of the diluted sample to each sample well.

Note: The sample dilution ratio is equivalent to 1:200.

4. Mix thoroughly, cover with a sealing film, and incubate at 37℃ (a water bath is recommended) in the dark for 30 minutes.

5. Discard the liquid from each well, add 350 μL of working wash solution to each well, let it sit for 30 seconds, then discard the solution. Repeat the washing step five times, and finally blot dry with clean absorbent paper.

6. Add 100 μL of enzyme-labeled reagent to each well (except the blank well). Cover with a sealing membrane and incubate at 37℃ in the dark for 30 minutes.

7. Wash, following Step 5.

8. Add 50 μL of substrate solution and 50 μL of developing solution to each well in sequence, mix thoroughly, cover with a sealing film, and incubate at 37℃ in the dark for 10 minutes.

9. Add 50 μL of stop solution to each well, mix thoroughly, and measure the absorbance (OD value) of each well at 450 nm (a reference wavelength of 630 nm may be used). If no reference wavelength is available, zero the instrument using the blank well (i.e., subtract the absorbance of the blank well from the absorbance of each well).

To determine how to interpret the test results, please contact our company!