Cattle and sheep brucellosis indirect enzyme-linked immunosorbent assay antibody detection kit and preparation method thereof

An enzyme-linked immunosorbent assay, Brucella bovis and sheep technology, applied in measuring devices, instruments, scientific instruments, etc., can solve the problems of low specificity and low sensitivity, and achieve good repeatability and specificity. The effect of good performance and good detection effect
CN103543261AInactive Publication Date: 2014-01-29INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS +1
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
Publication Date
2014-01-29
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention relates to a cattle and sheep brucellosis indirect enzyme-linked immunosorbent assay antibody detection kit and a preparation method thereof, belongs to the field of detection of pathogens of zoonotic infections and aims at solving the problems of low sensitivity, poor specificity and the like in detection of Brucella by adopting other antigens. The kit is assembled by taking a recombinant Brucella BCSP31 protein with expression induced by SUMO as an expression vector in Escherichia coli to serve as an antigen-coated ELISA (enzyme-linked immunosorbent assay) plate, adding serum to be detected and rabbit anti-bovine HRP-IgG or rabbit anti-goat HRP-IgG and adding a washing buffer solution, a blocking solution, a TMB (3,3',5,5'-Tetramethylbenzidine) primer solution and a stop solution for matching. The prokaryotic expression vector, namely the SUMO, is utilized to express the Brucella BCSP31 protein in an efficient and soluble manner, the kit is used for establishing a Brucella ELISA detection method, the detection has better repeatability and specificity, and cattle and sheep Brucella can be fast and efficiently detected by utilizing the method.
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Description

technical field

[0001] The invention relates to the technical field of detection of zoonotic infectious disease pathogens, in particular to an indirect enzyme-linked immunosorbent assay antibody detection kit for cattle and sheep brucellosis and a preparation method thereof. Background technique

[0002] The Brucella cell surface protein 31ku (BCSP31) gene was cloned, sequenced and expressed in vitro in 1988. It encodes a soluble cell surface protein with an immunogenicity of 31ku. Except for Brucella ovis epididymis, The expression of BCSP31 protein could be detected in the other 6 species of Brucella. The BCSP31 gene of Brucella has a homology of 99.3% in 8 Brucella strains from different genera. It is a Brucella species-specific gene and is highly conserved.

[0003] At present, some scholars have expressed BCSP31 protein by eukaryotic expression method, and used its eukaryotic expression product to immunize mice to study its ability to resist Brucella infection in mice;...

Claims

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