Competitive ELISA method based on foot-and-mouth disease A type VP1 protein and its monoclonal antibody

Abstract

The invention relates to a competitive ELISA method based on a foot-and-mouth disease A type VP1 protein and its monoclonal antibody, also relates to a preparation method of the foot-and-mouth disease A type VP1 protein, and a preparation method of the monoclonal antibody of the foot-and-mouth disease A type VP1 protein, and belongs to the technical field of animal immunological detection. In the invention, a primer pair C1 and C2 and a primer pair E1 and E2 are amplified to obtain a gene sequence of the foot-and-mouth disease A type VP1 protein, the foot-and-mouth disease A type VP1 protein is obtained by constructing an expression plasmid, introducing the expression plasmid into a prokaryotic expression host and carrying out inducible purification, the foot-and-mouth disease A type VP1 protein monoclonal antibody is obtained by treating the foot-and-mouth disease A type VP1 protein as an antigen through a hybridomas technology, and the competitive ELISA method used for detecting a foot-and-mouth disease A type antibody is established based on the foot-and-mouth disease A type VP1 protein and its monoclonal antibody. The detection method has a strong specificity and a good stability, and can be used for detecting a foot-and-mouth disease A type serum antibody. By comparing a result obtained through the detection method with a liquid phase blocking ELISA kit, the coincidence rate is 95.8%.

Description

technical field [0001] The invention belongs to the technical field of animal immunology detection, and in particular relates to a foot-and-mouth disease type A VP1 protein and a preparation method thereof, and a competition ELISA method based on the prepared foot-and-mouth disease type A VP1 protein and a monoclonal antibody thereof. Background technique [0002] Foot-and-mouth disease is a highly contagious and economically important animal disease caused by foot-and-mouth disease virus (FMDV), infecting cattle, pigs, sheep, goats and other artiodactyls. After animals are infected with foot-and-mouth disease, blisters will appear in the mouth, hooves, nipples and other parts to form erosions, and the diseased animals are generally benign. Although the death rate of foot-and-mouth disease is low, due to its high incidence rate of almost 100% and its rapid spread, it restricts the development of animal husbandry and the participation of countries in international trade of an...

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Problems solved by technology

[0006] The purpose of the present invention is to provide a competitive ELISA method based on the foot-and-mouth disease type A VP1 protein and its monoclonal antibody in view of the shortcomings of the existing detection technology for foot-and-mouth disease type A serum, such as high detection cost, troublesome detection operation, and large error in results. To the preparation of monoclonal antibody to type A foot-and-mouth disease protein VP1, and its application in the detection of pathogens and antibodies and the development of vaccines, etc.; established a monoclonal antibody competition ELISA method to detect type A antibody, and formed a type A foot-and-mouth disease immune monitoring technology platform

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