1. Purpose

        In the purification and prevention and control of brucellosis (hereinafter referred to as brucellosis), daily testing is required, and the technical teachers in charge of testing in the laboratory always have many questions about the new method. In the commercially available ELISA kits, people often doubt whether they are authentic and reliable, while the tiger red plate test and the test tube coagulation method according to the test protocol are time-consuming, sensitive, and difficult to judge. For this reason, our laboratory compares the sensitivities of the three tests for brucellosis in order to help others choose which method to use for the test.

        2. Materials and methods

        2.1 Materials:

        2.1.1 Lexenosis Competition Antibody Detection ELISA Kit (Lot Code: TE190701-1)

        2.1.2 Brucellosis tiger red plate agglutination antigen, purchased from China Institute of Veterinary Drug Control, batch number: 20190305.

        2.1.3 Brucellosis in vitro agglutination antigen: purchased from China Institute of Veterinary Drug Control, batch number: 20180607.

        2.2 Sample Selection

        2.2.1 Sample selection: The Animal Disease Prevention and Control Center of a city in Henan Province provided 90 samples of bovine and sheep serum, a total of 180 serum samples. All samples were tested by tiger red and ELISA kits, and the suspected positive and positive samples were tested by test-tube agglutination.

        2.2.2 The operation of the test process and the entry of the test results are carried out by different personnel, and the test results are sorted out and statistically analyzed to obtain the clinical research results, to ensure that all the contents of the research plan are strictly adhered to and the research data are filled in correctly, to prevent errors in the experimental data when recorded, and to ensure its completeness and accuracy.

3. Analysis of experimental results

        3.1 Analysis of kit test results:

        A total of 180 samples were clinically selected in the laboratory, all of which were determined using the Robosis competition ELISA detection kit produced by Luoyang Leprosheng Company, and the results of the determination are shown in Table 1 below:

Table 1 Results of the Competitive Testset for Leprosenbosis

Note: Samples marked in yellow are those that test positive.

From the results of the above table, it can be seen that the sheep serum used in the test was negative, and the bovine serum 3, 5, 7-9, 11, 13-21, 23-25, 27, 29, 36, 49 and 81, a total of 23 sera were tested positive.

2. The test results of the tiger red plate are as follows:

Figure 1: Results of a bovine blood sample

Figure 2: Test results of sheep blood samples

  As can be seen from Figure 1 and Figure 2 above, all sheep serum test results were negative, and a total of 8 samples of bovine serum 3, 5, 8, 14-16, 19 and 24 were tested positive.

        3. Test results of test tube agglutination of cloth disease:

        A total of 23 serum samples were tested for test-tube agglutination of the samples that were positive by both methods, and the serum samples were 3, 5, 7-9, 11, 13-21, 23-25, 27, 29, 36, 49 and 81, respectively. All serum samples were renumbered from smallest to largest, numbered A1-A23. A total of 4 positive samples were detected in this test, and the test results are shown in Figure 3 below:

As can be seen from the above picture, the four samples 3, 5, 14 and 15 of bovine serum were detected by the test-tube agglutination method of brucellosis, and the rest of the samples were negative. The overall results are analyzed in Table 2.

 4. Analysis and summary

        The results of the above three methods can be clearly seen that in the given 180 cattle and sheep samples, the positive detection rate of the cloth disease kit is 12.78%, and that of the tiger red plate is 4.44%, and the samples made by the test tube agglutination are all positive samples detected by the above two methods, so the positive rate of this method is lower, and the rough calculation is only 2.22%. Therefore, for the 180 serum samples sent for testing, the positive detection rate was: brucellosis kit> tiger red plate> test tube agglutination.

        Since the agglutination of tiger red plate and test tube is observed and interpreted by the human owner, the method has a great relationship with whether the operator has relevant experience and whether the operation is proficient, and the repeatability is relatively poor. However, the detection judgment of the kit relies on the microplate reader to determine the reading value, so the reproducibility of the method will be better. And in terms of methodology, the sensitivity of the kit is higher than that of the tiger red plate, and the sensitivity of the tiger red plate is higher than that of the test tube agglutination. Those who wish to carry out testing can choose the appropriate method according to their needs.