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Salmonella pullorum and mycoplasma synoviae double-plate agglutination antigen as well as preparation method and application thereof

A technology for Mycoplasma synovialum and Salmonella, applied to biochemical equipment and methods, methods based on microorganisms, bacteria, etc., to achieve good stability, rapid diagnosis, and cost-saving detection effects

Active Publication Date: 2021-08-17
SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology describes an improved way for diagnosing bacterial or parasitic infectious agents by combining salmolysin with certain substances that specifically target these organisms' pathogenesis. These compositions have several technical benefits such as being highly sensitive but quicker than traditional methods like enzyme immunoassay (EIA) which requires multiple steps before results become available. Additionally, this new technique allows for easier observation overall due to its ability to identify both types of microorganism simultanously during testing without causing any harmful side reactions from other materials used together.

Problems solved by technology

This patented technical problem addressed in this patents relates to prevention or treatment of diseases associated with muscle tissue such as rheunts (chloroquinoxylam) syndrome, septicemia, lymphatic system disorder, etc., particularly when treating young birds from broiler houses where they may pass into other areas during their lifespan.

Method used

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  • Salmonella pullorum and mycoplasma synoviae double-plate agglutination antigen as well as preparation method and application thereof
  • Salmonella pullorum and mycoplasma synoviae double-plate agglutination antigen as well as preparation method and application thereof
  • Salmonella pullorum and mycoplasma synoviae double-plate agglutination antigen as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1, Preparation of double plate agglutination antigen of Salmonella pullorum pullorum and Mycoplasma gallinarum

[0038] The double plate agglutination antigen of Salmonella pullorum and Mycoplasma gallinarum mentioned in this example is made of Salmonella pullorum C79-1 and Mycoplasma pullorum GX11-T, Salmonella pullorum strain C79-1 and chicken The Mycoplasma synoviae strains GX11-T are all preserved in the China Center for Type Culture Collection, and the preservation numbers are: CVCC79207 and CVCC2960, respectively.

[0039] Salmonella pullorum antigen was prepared by the following method:

[0040](1) Preparation of seed bacteria liquid: after recovery and propagation of Salmonella pullorum C79-1 strain in TSB liquid medium, inoculate TSA solid plate medium, culture at 37°C for 18 hours, select typical colonies and inoculate TSA solid slant medium, 37 After cultivating at ℃ for 20 hours, wash the bacterial lawn with Martin's liquid medium, expand the cultur...

Embodiment 2

[0048] Example 2, the method of using double plate agglutination antigen of Salmonella pullorum and Mycoplasma gallinarum

[0049] 30 minutes before use, take out the agglutinated antigen, negative or positive serum and the serum to be tested from the double plate of Salmonella pullorum and Mycoplasma gallinarum from the refrigerator, and let it reach room temperature. Use a pipette to absorb a drop (0.025mL-0.05mL) of the well-mixed diagnostic antigen, drop it vertically on a glass plate, and then quickly add a drop (0.025mL-0.05mL) of the same amount of serum to be tested next to the antigen. Mix the serum and antigen evenly, spread it into a sheet with a diameter of 1cm-2cm, shake the glass plate constantly, and observe the results within 2min. The test should be carried out under the condition of 20℃~25℃. Each batch of tests should be set positive serum and negative serum control.

[0050] Judgment of the results: if more than 50% (++) of agglutination occurs within 2 mi...

Embodiment 3

[0054] Example 3, double plate agglutination antigen specificity experiment of Salmonella pullorum and Mycoplasma gallinarum

[0055] The prepared double-plate agglutinated antigens of Salmonella pullorum and Mycoplasma gallinarum were mixed with Salmonella gallinarum positive serum, Mycoplasma gallinarum positive serum, Escherichia coli positive serum, Staphylococcus gallinarum positive blood, Haemophilus paragallinarum positive serum , chicken Newcastle disease positive serum, avian influenza positive serum, chicken infectious bronchitis positive serum, chicken adenovirus positive serum, etc. were subjected to plate agglutination reaction, and the results were as follows image 3 As shown, only the positive serum of Salmonella gallinarum and the positive serum of Mycoplasma synovialum had agglutination reaction, and the others were all negative.

[0056] Example 3, clinical application of double plate agglutination antigen of Salmonella pullorum and Mycoplasma gallinarum

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Abstract

The invention discloses a salmonella pullorum and mycoplasma synoviae double-plate agglutination antigen as well as a preparation method and application thereof, and the preparation method comprises the following steps: respectively carrying out resuscitation passage on salmonella pullorum C79-1 and mycoplasma synoviae GX11-T to prepare seed bacterial liquid, inoculating the seed bacterial liquid into a corresponding culture medium, carrying out enlarged culture, harvesting, respectively inactivating by a formaldehyde solution, concentrating, mixing the two bacterial liquids according to a certain volume ratio, adding a crystal violet solution with the final concentration of 3% by mass fraction for dyeing and glycerol with the volume fraction of 10%, uniformly mixing, and subpackaging, thereby obtaining the product. The technological method is simple and reasonable, low in cost and good in stability, the prepared double-plate agglutination antigen can detect two epidemic disease antibodies at the same time through one-time reaction, the advantages of being high in sensitivity, rapid in diagnosis, high in specificity, easy to observe the agglutination effect and the like are achieved, the detection time and the detection cost are remarkably saved, and the detection efficiency is improved. The product can be clinically used for rapidly detecting the two antibodies of the pullorum disease and the mycoplasma synoviae, diseased chickens are eliminated, and disease purification is achieved.

Description

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Claims

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Application Information

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Owner SHANDONG BINZHOU ANIMAL SCI & VETERINARY MEDICINE ACADEMY
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