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Germ model screening system and screening method

A model and screening technology, applied in biological systems, analytical materials, biological material analysis, etc.

Pending Publication Date: 2021-04-06
CHONGQING ACAD OF ANIMAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no relevant literature on effective vaccines for mixed infection of Cryptobacterium pyogenes, Corynebacterium pseudotuberculosis, Staphylococcus aureus, Proteus mirabilis and other pathogenic bacteria

Method used

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  • Germ model screening system and screening method
  • Germ model screening system and screening method
  • Germ model screening system and screening method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Embodiment 1 screening system

[0106] refer to figure 1 , a system for screening infectious bacteria models, including microplates, heaters, reagents, databases, microplate readers, and data comparison output modules;

[0107] Among them, the enzyme label plate includes polystyrene / polyvinyl chloride material standard plate, including 96 holes or 48 holes;

[0108] Among them, the reagents include antigen coating solution, enzyme-labeled antibody, chromogenic solution, bacterial infection model serum or immune model serum;

[0109] Among them, the database produces the absorbance of different antibody amounts for healthy or infected or immune animal models, which can be imported and deleted;

[0110] Wherein, the microplate reader is set to have a wavelength of 450nm or 410nm.

[0111] Wherein, the chromogenic solution is TMB single-component chromogenic solution.

[0112] Wherein, the antigen coating solution includes PLO coating solution with a concentration of 0...

Embodiment 2

[0113] Example 2 Bacterial Culture and Preparation of Recombinant Protein

[0114] (1) Bacterial culture

[0115] Inoculate a single colony of Cryptobacterium pyogenes (biological deposit number: CCTCC NO: M2014037, GenBank NO: CP012649) in TSB medium containing 8% calf serum, and shake at 37°C for 48 hours;

[0116] A single colony of Corynebacterium pseudotuberculosis was inoculated in TSB medium, and shaken at 37°C for 24 hours;

[0117] Single colonies of Staphylococcus aureus, Proteus mirabilis, and Escherichia coli were inoculated in LB medium respectively, and cultured with shaking at 37°C overnight.

[0118] (2) Identification of Cryptobacter pyogenes PLO

[0119] The C. pyogenes freshly cultured in Example 1 was centrifuged at 6 000 r / min for 10 min, and the supernatant was retained. The supernatant was filtered through a 0.22 μm filter membrane and concentrated by ultrafiltration with a molecular cut-off of 10,000ku. The PLO in the concentrated solution was ident...

Embodiment 3

[0135] Embodiment 3 test analysis

[0136] (1) Cross immune protection test

[0137] The inactivated vaccine of C. pyogenes made with aluminum gel adjuvant was used to immunize mice according to the method of step (4) in Example 2, and a total of 45 mice were immunized. Two weeks after the third immunization, the immunized mice were intraperitoneally inoculated with a lethal dose of Corynebacterium pseudotuberculosis, and 15 unimmunized healthy mice were inoculated intraperitoneally with a lethal dose of Corynebacterium pseudotuberculosis. The death rate of the mice was counted.

[0138] Two weeks after the third inactivated C. pyogenes vaccine, a lethal dose of Corynebacterium pseudotuberculosis was inoculated intraperitoneally. The survival rate of the immunized mice was 53.3% (24 / 45), and one unimmunized mouse survived. This indicated that inactivated C. pyogenes immunization had a cross-immune protective effect on Corynebacterium pseudotuberculosis.

[0139] (2) Agglutina...

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Abstract

The invention belongs to the field of germ screening and particularly relates to a system and a method for screening a model infected with arcanobacterium pyogenes or corynebacterium pseudotuberculosis. The system comprises an elisa plate, a heater, a reagent, a database, an elisa instrument and a data comparison output module, wherein the elisa plate comprises a polystyrene / polyvinyl chloride material label plate which comprises 96 holes or 48 holes; the reagent comprises an antigen coating solution, an enzyme-labeled antibody, a developing solution and a germ infection model or immune model serum; the database generates absorbance of different antibody amounts for a healthy or infected or immunized animal model, and the absorbance can be imported and deleted; and the wavelength of the microplate reader is set to be 450 nm. The screening system provided by the invention is complete in database, accurate in screening result, capable of quickly screening out the types of the bacteria infecting the bacteria model, simple and convenient.

Description

technical field [0001] The invention belongs to the field of germ screening, and in particular relates to a system and method for screening models infected with Cryptobacterium pyogenes or infected with Corynebacterium pseudotuberculosis. Background technique [0002] Trueperella pyogenes infection occurs in domestic and wild animals all over the world, and infection usually leads to suppurative pneumonia, mastitis, endometritis, abortion, stillbirth and other symptoms in a variety of animals. Goats infected with C. pyogenes mainly have clinical symptoms such as weight loss, superficial lymphadenitis, and suppurative pneumonia. Dissection shows purulent foci in the lungs and other organs, showing widening of the alveolar septum, thickening of the alveolar wall, pulmonary fibrosis, and multiple Pathological changes such as persistent infiltration of inflammatory cells in organs, suppurative inflammation, etc. Cryptobacterium pyogenes is a part of the microbial flora of anima...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/58G01N33/68G16B5/00G16B50/30
CPCG01N33/56911G01N33/6854G01N33/581G16B5/00G16B50/30G01N2469/20
Inventor 沈克飞许国洋付利芝闫志强张素辉杨柳徐登峰
Owner CHONGQING ACAD OF ANIMAL SCI
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