A detection test paper card for detecting Brucella bovis antigen by sandwich method
A technology for detecting Brucella bovis and test strips, which is applied in the field of immunological detection, can solve the problems of high false positive rate, lack of technical personnel, long reaction time, etc., achieve high sensitivity and repeat rate, easy to observe the difference, and good stability Effect
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Embodiment 1
[0034] Embodiment 1 Utilizes the sandwich method to detect the detection test paper card structure of the Brucella bovis antigen
[0035] Detection test paper card for detecting Brucella bovis antigen by sandwich method, such as figure 1 and figure 2 As shown, the detection test paper card is composed of a casing 1 and a test strip 2 inside it, and the casing 1 is composed of an upper plate 3 and a lower plate 4, the upper plate 3 has a detection window 5 and a sample injection hole 6, and the test strip 2 It consists of a bottom plate 7 and a sample absorption pad 8 , a colloidal gold marker pad 9 , and a detection reaction area 10 pasted on the bottom plate 7 in sequence. Colloidal gold marker pad 9 is coated with Brucella antibody and colloidal gold marker. The detection reaction area 10 is composed of a test area 11 coated with a Brucella antibody-carrier protein conjugate and a quality control area 12 coated with a Brucella antigen.
[0036] The carrier protein is bov...
Embodiment 2
[0039] Embodiment 2 detects the method for bovine brucella antigen
[0040] 1. Preparation of Brucella bovis antibody-colloidal gold marker
[0041] (1) Dialyze the Brucella bovis antibody in 0.005M / L pH7.0 NaCl solution overnight at 4°C, centrifuge at 10000-12000rpm for 1h at 4°C;
[0042] (2) the pH of the colloidal gold solution of OD526 is adjusted to 4.0-9.0;
[0043] (3) The Brucella bovis antibody obtained by centrifugation in step (1) is diluted to 5 μg / ml~50 μg / ml with a borate buffer solution of 0.005M / LpH9.0, and 1ml of the diluted Brucella bovis is taken The antibody was added to 1ml of colloidal gold solution, oscillated and mixed, and left to stand for 5 minutes, then 0.1ml of 10% NaCl solution was added, mixed and left to stand for 2 hours, and centrifuged to obtain the colloidal gold-labeled Brucella bovis antibody.
[0044] 2. Preparation of Brucella bovis antibody-carrier protein conjugate
[0045](1) Dissolve 5 mg of Brucella bovis antibody and 2.5 mg of ...
Embodiment 3
[0057] Embodiment 3 detects the test strip false positive rate and the false negative test experiment of bovine brucella antigen
[0058] 1. False Positive Tests
[0059] (1) Take 100 parts of common bovine blood samples, and use a blood pretreatment device to process the blood samples;
[0060] (2) Add 100 μL each of 100 processed common bovine blood samples and 100 μL deionized water into the sample injection hole;
[0061] (2) After the blood sample and deionized water flow through the test area and quality control area of the respective test strips, determine whether the sample contains Brucella bovis antibody. The determination rules are as follows:
[0062] Negative (-): Only a purple-red band appears in the quality control area, and no purple-red band appears in the test area;
[0063] Positive (+): Two purple-red bands appear, one in the test area and the other in the quality control area;
[0064] Invalid: When the quality control area does not display a purple-r...
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