Double antibody sandwich ELISA kit used for Seneca viral antigen detection and application thereof
A technology of antigen detection and double-antibody sandwich, which is applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of complex operation steps and the inability to realize real-time detection of a large number of samples, and achieve the effect of avoiding cross-reaction and shortening the detection time
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Embodiment 1
[0056] Example 1 Establishment of a double-antibody sandwich ELISA detection method for Seneca virus antigen detection
[0057] 1. Materials and methods
[0058] 1.1 Materials
[0059] 1.1.1 Experimental animals and antigens
[0060] 1.5-2.0 kg healthy male rabbits were purchased from the Lanzhou Veterinary Research Institute of the Chinese Academy of Agricultural Sciences; the inactivated antigen of porcine Seneca Valley virus (SVV) was prepared and provided by the Lanzhou Veterinary Research Institute of the Chinese Academy of Agricultural Sciences.
[0061] 1.1.2 Main reagents
[0062] 96-well microtiter plates were purchased from Costar Company, 2mol / L H 2 SO 4 Provided by the detection team of Lanzhou Veterinary Research Institute, fetal bovine serum was purchased from GIBICO, TMB was purchased from SurModics, carbonate buffer capsules were purchased from Sigma, and unstained protein markers were purchased from Fermentas.
[0063] 1.2 Method
[0064] 1.2.1 Preparati...
Embodiment 2
[0141] The assembling and using method of embodiment 2 kit
[0142] 1. Assembly of kit
[0143] (1) ELISA plate: Seneca virus guinea pig anti-IgG-coated ELISA plate, prepared according to the method in Example 1, and stored in a refrigerator at 4°C.
[0144] (2) 10× washing solution: the 10× washing solution contains 5v / v% Tween-20, 8w / w% sodium chloride, 0.2w / w% potassium chloride, 2.9w / w% disodium hydrogen phosphate and 0.2w / w% potassium dihydrogen phosphate aqueous solution, pH7.4;
[0145] (3) 10× sample diluent: the 10× sample diluent contains 5w / w% bovine serum albumin, 5v / v% Tween-20, 8w / w% sodium chloride, 0.2w / w% chloride An aqueous solution of potassium, 2.9w / w% disodium hydrogen phosphate and 0.2w / w% potassium dihydrogen phosphate, pH7.4;
[0146] (4) Enzyme-labeled antibody diluent: 0.01mol / L PBS buffer containing 1v / v% glycerol, 0.5w / v%BSA, 1w / v% casein and 0.05w / v%Procline300, pH7.4, according to Example 1 preparation.
[0147] (5) Enzyme-labeled antibody wo...
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