Solid phase competition ELISA kit for detecting Seneca valley virus antibody, and applications thereof,
An antibody detection and solid-phase competition technology, applied in biological testing, measuring devices, material inspection products, etc., can solve the problems of complex operation steps, inability to realize real-time detection of a large number of samples, etc., to avoid cross-reaction and shorten the detection time. Effect
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Embodiment 1
[0052] Example 1 Establishment of a solid-phase competitive ELISA detection method for Seneca virus antibody detection
[0053] 1. Materials and methods
[0054] 1.1 Materials
[0055] 1.1.1 Experimental animals and antigens
[0056] 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.
[0057] 1.1.2 Main reagents
[0058] 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.
[0059] 1.2 Method
[0060] 1.2.1 Preparati...
Embodiment 2
[0132] The assembling and using method of embodiment 2 kit
[0133] 1. Assembly of kit
[0134] (1) ELISA plate: an ELISA plate coated with Seneca virus inactivated antigen, prepared according to the method in Example 1, and stored in a refrigerator at 4°C.
[0135] (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;
[0136] (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, pH 7.4;
[0137] (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.
[0138] (5) Enzyme-labeled re...
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