Immunoassay kit by using light initiated chemiluminescent assay for bovine mycoplasma antibody and detection method thereof

Abstract

The invention discloses an immunoassay kit by using a light-initiated chemiluminescent assay for a bovine mycoplasma antibody and a detection method thereof. The kit comprises bovine mycoplasma antibody P30 protein monoclonal antibodies, goat anti-mouse IgG conjugated acceptor microbeads, His-tagged bovine mycoplasma antibody P30 protein antigens, and nickel chelated donor magnetic beads. The invention prepares outer membrane protein of bovine mycoplasma antibody P30 by using the genetic engineering technology, prepares bovine mycoplasma antibody P30 protein monoclonal antibodies by using thePEG1500 cell fusion technology, and the purified recombinant P30 protein is used as a product of interaction between the antigen and the prepared monoclonal antibody, competing with the monoclonal antibody by the antibody in the sample to bind and recombine the P30 protein, and establishing a method for detecting competitive bovine mycoplasma antibody AlphaLISA. According to the immunoassay kit byusing light initiated chemiluminescent assay for the bovine mycoplasma antibody and the detection method thereof, the detection method has the advantages of short detection time, good specificity, good sensitivity, no need for washing, simple operation and low detection cost.

Description

technical field [0001] The invention relates to the fields of veterinary medicine, immunology and biological products, in particular to a photo-activated chemiluminescence immunoassay kit for mycoplasma bovis antibody and a detection method thereof. Background technique [0002] Mycoplasma bovis (Mb) is the smallest and simplest prokaryon that can replicate autonomously. It can cause bovine pneumonia, mastitis, arthritis and other diseases, causing huge economic losses and seriously threatening the healthy development of my country's cattle industry. . At present, the commonly used detection methods for Mycoplasma bovis in veterinary laboratories include biological detection, molecular biological detection and immunological detection. Biological detection is mainly through the traditional isolation and identification of pathogens. Because mycoplasma is a facultative anaerobic organism, it has extremely strict nutritional requirements, and the culture period is long (it takes...

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Problems solved by technology

[0004] Aiming at the deficiencies of the above-mentioned prior art, the object of the present invention is to provide a Mycoplasma bovis antibody photoactivated chemiluminescence immunoassay kit and its detection method to solve the problems of cumbersome operation and poor specificity in the existing Mycoplasma bovis detection method

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