Method for detecting bovine brucellosis by competence ELISA (Enzyme-Linked Immunosorbent Assay) kit

Abstract

The invention discloses a method for detecting bovine brucellosis by a competence ELISA (Enzyme-Linked Immunosorbent Assay) kit, belonging to the field of biotechnical detection. The method comprises the steps: coating bovine brucella A99 subjected to ultrasonic splitting by an ELISA enzyme-labeled plate; then adding an anti-brucella specific monoclonal antibody bru-5C10 and a diluted to-be-detected serum sample in plate holes of the ELISA coated plate; and simultaneously respectively adding the anti-brucella specific monoclonal antibody bru-5C10 and known negative and positive serum in the plate holes of the ELISA enzyme-labeled plate, washing with PBST (polybutylene terephthalate) after water bathing, respectively adding an anti-mouse IgG (Intravenous Gamma Globulin) enzyme-labeled antibody in each plate hole, performing a chromogenic reaction after incubation and washing, measuring an OD450 value of each plate hole, and calculating an inhibition rate of each plate. According to the method, yersinia enterocolitica O9 and escherichia coli O157 high-immunity positive serum are detected by using an established cELISA method, a PI (inhibition rate) value is lower than 30 percent, and the generation of a false positive result can be effectively eliminated.

Description

technical field [0001] The invention belongs to the field of biotechnology detection, in particular to the technical field of detection of brucellosis in cattle and sheep. Background technique [0002] Brucellosis (brucellosis) is an important zoonotic disease caused by Brucella, and ruminants, especially cattle and sheep, are susceptible to infection. According to the antigen type and host type, Brucella can be divided into 6 types. The general symptom of sick female animals is abortion, while males can cause reproductive disorders such as infertility. In recent years, brucellosis has shown a rebound trend worldwide. In China, the damage caused by B. abortus and B. melitensis is particularly extensive and serious. The disease has caused huge economic losses to animal husbandry and public health, and it is of great practical significance to strengthen the prevention and control of brucellosis. [0003] Accurate diagnosis of brucellosis is the primary measure to prevent the...

AI Technical Summary

Problems solved by technology

[0004] So far, monoclonal antibodies against the A antigen, M antigen, C antigen and C / Y antigen located on the lipopolysaccharide O chain (O-LPS) have been prepared; on the other hand, scholars have found that some specific outer membrane proteins There is also immunogenicity, such as OMP31 and other cELISA methods established using these monoclonal antibodies have long been used in the serological detection of brucellosis, but due to the limitations of the sensitivity and specificity of different monoclonal antibodies, as well as the interference of cross-reactions, in In the actual detection, there are often missed and wrong detection results.

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