Detection reagent for mycoplasma bovis antibody and preparation method thereof

A technology for detection of mycoplasma bovis and antibodies, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, biological testing, etc., can solve the problems of unpublished, lack of epidemiological information, complicated operation steps, etc., and avoid storage time Short, easy to store for a long time, high hemagglutination titer effect

CN104062439AActive Publication Date: 2014-09-24LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
Publication Date
2014-09-24

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Abstract

The invention relates to a biological detection reagent and a preparation method thereof, and particularly relates to a detection reagent for a mycoplasma bovis antibody and a preparation method thereof. The detection reagent for the mycoplasma bovis antibody is composed of one part of a mycoplasma bovis P48 recombined and expressed fusion protein antigen and one part of 10% hydroformylation-tanned sheep red blood cells (v / v) which are uniformly mixed. The gene sequence of the P48 recombined and expressed fusion protein antigen for the detection reagent for the mycoplasma bovis antibody is SEQ ID No.1. The detection reagent can be used for really detecting mycoplasma bovis and has the advantages of long preservation time, high sensitization titer, easiness in long-time preservation, high blood coagulation titer, simplicity in operation, convenience and rapidness, low cost, safety and stability, no toxin, no environmental pollution and the like.
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Description

technical field

[0001] The invention relates to a biological detection reagent and a preparation method of the detection reagent. Specifically, the invention relates to a detection reagent for mycoplasma bovis antibody and a preparation method thereof. Background technique

[0002] Mycoplasma bovis is an important pathogenic pathogen that infects cattle. In 1961, Hale first isolated the pathogen from cow's milk with mastitis in the United States. In 1976, it was first reported to be related to bovine respiratory diseases. It has been confirmed that Mycoplasma bovis can cause other diseases such as arthritis, keratoconjunctivitis, otitis, reproductive tract inflammation, miscarriage and infertility in addition to bovine pneumonia and mastitis. Since 2008, the outbreak of "contagious mycoplasma bovis pneumonia" characterized by necrotizing pneumonia has occurred in some areas of my country newly imported beef cattle from other places. It has caused huge economic losses to my c...

Examples

Embodiment Construction

[0014] The present invention is explained in detail below in conjunction with examples.

[0015] 1. Preparation of recombinantly expressed fusion protein antigen of Mycoplasma bovis P48

[0016] a. Synthesis of p48 gene of Mycoplasma bovis and identification of recombinant plasmid

[0017] The codons of the published Mycoplasma bovis p48 gene (Genebank: NC_014760.1) were optimized according to the preference of codons in Escherichia coli, and four TGAs expressing tryptophan (UGA ) was optimized to TGG (UGG), because this codon is a stop codon in Escherichia coli, and at the same time, restriction sites BamH I and Xho I were added at both ends, and the optimized gene sequence is SEQ ID № 1 ;

[0018] The optimized p48 gene sequence was synthesized into the pet32a expression vector plasmid by Invitrogen Company. After the synthesis, it was identified by sequencing and enzyme digestion, and the positive recombinant plasmid after successful identification was named Pet32-a(+)-p4...