Test paper card for detecting swine transmissible gastroenteritis virus antibody and preparation and detection method of test paper card
A technology for infectious and gastroenteritis, applied in the field of immunological detection, can solve the problems of large error, low sensitivity, poor stability, etc., and achieve the effect of shortening time, high sensitivity and good stability
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Embodiment 1
[0024] Embodiment 1, the preparation of test paper card
[0025] 1. Preparation of fluorescent microsphere-antibody complexes
[0026] (1) Cleaning: Take 50μl of fluorescent microspheres into a 1.5ml centrifuge tube, add 1ml of 0.01M MES buffer, shake and mix, centrifuge at 15000r / min for 15min, discard the supernatant, add 1ml of 0.01M MES buffer, and ultrasonically beat Loose microspheres; repeat this step three times, the purpose of cleaning the microspheres has been achieved, and finally the microsphere liquid after cleaning is obtained;
[0027] (2) Activation: add 250μl EDC solution to 1ml of microsphere solution, activate in the dark for 2h, centrifuge at 15000r / min for 15min, discard the supernatant, add 1ml of 0.01M MES buffer, ultrasonically break up the microspheres, and centrifuge at 15000r / min 15min, discard the supernatant;
[0028](3) Labeling: add 1ml of 0.05M borate buffer, disperse by ultrasonic, add 20μg anti-E2 protein monoclonal antibody and mix well, an...
Embodiment 2
[0048] Embodiment 2, detection method and comparative test of porcine transmissible gastroenteritis virus antibody in pig anal swab
[0049] 1. Wet the cotton swab with physiological saline and wipe the pig feces and rectum. Immediately insert the cotton swab into the sample tube containing the diluent, fully stir the cotton swab until all the samples on the cotton swab are dissolved in the diluent, and squeeze the cotton swab on the tube wall. Discard after drying, and the sample solution is left to stand for later use; the diluent is PBS with pH 7.4 and 0.05mol / L;
[0050] 2. Tear open the aluminum foil packaging bag of the test card, take out the test card, and place it on a flat and clean table;
[0051] 3. Take the prepared clarified sample supernatant with the matching straw, and drop 2-3 drops (about 60 μl) vertically and slowly into the sample well;
[0052] 4. Read the result in 5-10 minutes, insert the test paper card into the card slot of the fluorescence immunoass...
Embodiment 3
[0056] Embodiment 3, colloidal gold-labeled anti-E2 protein monoclonal antibody and the comparison test of the stability of fluorescent microsphere-labeled anti-E2 protein monoclonal antibody
[0057] 1. Place the prepared colloidal gold-antibody complex binding pad and the fluorescent microsphere-antibody complex binding pad in a blast drying oven at 37°C;
[0058] 2. On the 3rd day, 5th day, 1 week, 2 weeks, 4 weeks, 8 weeks, 12 weeks, 20 weeks, 36 weeks, and 48 weeks, test the negative samples and positive samples respectively, and observe their stability;
[0059] 3. The results are shown in Table 1 below.
[0060]
[0061] It can be seen from the above table 1 that the detection effect of the colloidal gold-labeled antibody complex decreases with the prolongation of the destruction test time, and the detection situation changes significantly at the 20th week; while the test strip using the fluorescent microsphere-antibody complex binding pad The positive samples showe...
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