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ELISA kit for rapidly detecting sheep mycoplasma pneumoniae antibody

A technology of Mycoplasma pneumoniae and a kit, which is applied in biological tests, measuring devices, material inspection products, etc., can solve the problems of less cross-reaction, difficulty, and time-consuming, and achieve the effect of strong specificity and high sensitivity

Active Publication Date: 2019-12-20
GUANGXI VETERINARY RES INST
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Problems solved by technology

The isolation and cultivation of the pathogen of sheep mycoplasma pneumonia is cumbersome, difficult, and time-consuming. Indirect hemagglutination detection methods have serious cross-reactions and poor repeatability. Molecular biological diagnostic methods such as PCR are expensive and should not be extended to the grassroots. However, the ELISA method is easy to operate and cross-reactive. Less response, good repeatability, and most of the places have been equipped with related equipment

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  • ELISA kit for rapidly detecting sheep mycoplasma pneumoniae antibody
  • ELISA kit for rapidly detecting sheep mycoplasma pneumoniae antibody
  • ELISA kit for rapidly detecting sheep mycoplasma pneumoniae antibody

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Embodiment Construction

[0026] The following will describe in detail the research process of the expression of the outer membrane protein of Mycoplasma ovis pneumoniae in the present invention, the identification of immunogenicity and the establishment of the ELISA detection method. Among them, Mycoplasma ovipneumoniae, positive and negative sera are prepared and stored by our laboratory; skimmed milk powder Difco TM Skim milk was purchased from Solarbio; enzyme-labeled secondary antibody: goat anti-mouse-Ig G was purchased from Zhongshan Jinqiao; other reagents were all domestically pure products.

[0027] 1. Preparation of the coating antigen. Use restriction enzymes to cut the MOL gene separately (the PCR amplification results are as follows: figure 1 ) Fragment and pET-32a expression vector, the digested product is recovered by gel and then ligated with T4 enzyme. The ligation reaction system is: MOL gene fragment 3uL, pET-32a vector 1uL, T4 ligase 1uL, water 5uL. The ligation was placed at 4°C ov...

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Abstract

The invention discloses an ELISA kit for rapidly detecting sheep mycoplasma pneumoniae antibody. The ELISA kit comprises a coated enzyme-labeled plate, negative standard serum, positive standard serum, an enzyme-labeled second antibody, a washing liquid, a diluents, a substrate liquid and an ending liquid. Recombination sheep mycoplasma pneumoniae in-vitro membrane protein MOL with relatively highimmunogenicity is acquired by building an expression carrier and is used as a coated antigen, and sheep mycoplasma pneumoniae antibody ELISA detection method and kit which are high in specificity andhigh sensitivity are built, the kit is used for sheep serum clinical sample detection, immunoprophylaxis and immunovalue study of sheep mycoplasma pneumoniae are favorably deepen, and a necessary technical means is provided for rapid detection of the sheep mycoplasma pneumoniae antibody.

Description

Technical field [0001] The invention relates to the technical field of antibody detection of Mycoplasma ovipneumoniae, in particular to an ELISA kit for rapid detection of antibodies to Mycoplasma ovipneumoniae. Background technique [0002] The research of recombinant expression in bacterial viruses is mainly to express the virulence gene, which can improve the immunogenicity of the target protein; when it is made into a vaccine immunization, the animal can produce antibodies with a higher protection rate, thereby effectively defending against pathogens Invasion. [0003] Mycoplasma sheep pneumonia, also known as infectious pleuropneumonia, is mainly composed of Mycoplasma ovipneumoniae (MO), Mycoplasma mycoplasma subspecies goat (MMC), Mycoplasma caprae subspecies goat pneumonia (MCC) and Mycoplasma capra goat subspecies (MCSC), etc. An infectious disease of sheep or goats caused by Mycoplasma species. Mycoplasma pneumonia has been reported in many provinces and regions in my c...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6854
Inventor 马春霞李军潘艳黎铭冯世文陶立彭昊李常挺钟舒红贺会利胡帅韦志锋兰美益
Owner GUANGXI VETERINARY RES INST
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