An indirect ELISA kit for detecting antibodies against Haemophilus parasuis

Abstract

The invention discloses an indirect ELISA (enzyme-linked immuno sorbent assay) kit for detecting a haemophilus parasuis antibody. The kit consists of an ELISA coating plate with haemophilus parasuis cytolethal distending toxin)-C protein serving as a coating antigen, a to-be-detected sample dilution plate, a positive contrast serum, a negative contrast serum, 20-time concentrated washing liquid, a serum sample dilution solution, an enzyme-labeled antibody working solution, a developing solution and a terminating solution. A judgment standard is that if an S / P value is less than 0.200, a sample to be detected is negative; if the S / P value is greater than or equal to 0.200, the to-be-detected sample is positive; the S / P value is obtained according to a formula: S / P value=(the mean value of the to-be-detected sample OD450nm-the mean value of a negative contrast OD450nm) / (the mean value of a positive sample OD450nm-the mean value of the negative contrast OD450nm). Due to the specificity test, the sensitivity test, the repetitiveness test, the coincidence rate test, the test for comparing the kit disclosed by the invention with a kit on sale, the clinical application test and the like, the kit disclosed by the invention has the characteristics of high specificity, high sensitivity, high repetitiveness and the like and is high in coincidence rate to the same type of products on sale home and abroad; the indirect ELISA kit can be used for clinical large-scale detection and epidemiological investigation for the haemophilus parasuis antibodies.

Description

technical field [0001] The invention relates to the technical fields of molecular biology, microbiology and animal infectious disease detection, in particular to an indirect ELISA kit for detecting antibodies to Haemophilus parasuis, and belongs to the field of prevention and control of Haemophilus parasuis disease. Background technique [0002] Haemophilus parasuis disease is an infectious disease caused by Haemophilus parasuis. Haemophilus parasuis ( H. parasuis ) is an NAD-dependent, non-motile, Gram-negative bacillus belonging to the genus Haemophilus of the Pasteurellaceae family. The bacterium is a commensal bacterium in the upper respiratory tract of pigs, which can invade the body under certain conditions and cause a serious systemic disease—Glasser's Disease, which is clinically manifested as multiple serosa in pigs arthritis, arthritis and meningitis. H. parasuis Susceptible to young pigs aged 2 weeks to 4 months, the disease mainly occurs before and after weani...

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Problems solved by technology

However, Zhao Qian (2010) showed that the gene sequence of outer membrane protein P5 in the 15 serotypes of H. The middle homology is high and conservation is good, while the homology of the outer membrane protein P2 is low, and some serotype gene sequences have segmental base deletions and multiple point mutations, which may affect its versatility as an envelope protein

Zhang Bin (2013) and others showed that the expressed outer membrane protein P5 protein can react with the positive sera of 15 serotype reference strains of Haemophilus parasuis by immunoblotting. The outer membrane protein P5 protein has good versatility, but indirect ELISA detection The results showed that the ELISA plates expressing the outer membrane protein P5 protein can cross-react with the positive sera of bacteria such as Pasteurella and Actinobacillus pleurodes, and the protein specificity is poor, which may cause false positive results

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