An ELISA kit for rapid detection of antibodies against Mycoplasma ovine pneumonia

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: 2022-07-22
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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  • An ELISA kit for rapid detection of antibodies against Mycoplasma ovine pneumonia
  • An ELISA kit for rapid detection of antibodies against Mycoplasma ovine pneumonia
  • An ELISA kit for rapid detection of antibodies against Mycoplasma ovine pneumonia

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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 sheep pneumoniae, the identification of immunogenicity and the establishment of the ELISA detection method in the present invention. Among them, Mycoplasma ovine pneumonia, positive and negative sera were prepared and preserved by our laboratory; skim milk powder Difco TM Skim milk was purchased from Solarbio; enzyme-labeled secondary antibody: goat anti-mouse-IgG was purchased from Zhongshan Golden Bridge Company; other reagents were domestic analytically pure products.

[0027] 1. The preparation of the coated antigen uses restriction endonucleases to digest the MOL gene respectively (the PCR amplification results are as follows: figure 1 ) fragment and pET-32a expression vector, the digested product was 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, wate...

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Abstract

The invention discloses an ELISA kit for rapid detection of Mycoplasma sheep pneumoniae antibody, comprising a coated enzyme labeling plate, negative standard serum, positive standard serum, enzyme labelled secondary antibody, washing solution, diluent, substrate solution and stop solution. By constructing an expression vector, a highly immunogenic recombinant Mycoplasma ovine pneumonia outer membrane protein MOL was obtained, which was used as a coating antigen, and a specific and highly sensitive ELISA detection method and kit for Mycoplasma ovine pneumoniae antibodies were established. The cassette is used for the detection of clinical samples of sheep serum, which will help to carry out in-depth research on the immune prevention and immune value of M. ovine pneumonia, and provide the necessary technical means for the rapid detection of M. ovine pneumonia antibodies.

Description

technical field [0001] The invention relates to the technical field of Mycoplasma ovine pneumoniae antibody detection, in particular to an ELISA kit for rapid detection of Mycoplasma ovine pneumoniae antibodies. Background technique [0002] The research of recombinant expression in bacterial viruses mainly focuses on recombinant expression of virulence genes, which can improve the immunogenicity of the target protein; making it into a vaccine for immunization can allow animals to produce antibodies with a higher protection rate, thereby effectively defending against pathogens invasion. [0003] Mycoplasma sheep pneumonia, also known as sheep infectious pleuropneumonia, is mainly caused by Mycoplasma ovine pneumonia (MO), Mycoplasma capricolum subspecies (MMC), Mycoplasma capricolum subspecies (MCC) and Mycoplasma capricolum subspecies (MCSC) An infectious disease of sheep or goats caused by Mycoplasma species. Mycoplasma pneumonia has been reported in many provinces and r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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