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Selective culture medium for detecting antigen bacterium content of swine fever/swine erysipelas/swine plague triple live vaccine, and preparation method and application thereof

A swine pneumonia, selective technology, applied in microorganism-based methods, biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of swine erysipelas antigen monitoring, etc., to solve the long-term problems of quality monitoring, accurate High precision and good repeatability

Inactive Publication Date: 2017-05-31
WENS FOOD GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is well known that the most fundamental determinant of the immune effect of a vaccine is the content of the effective antigen in the vaccine, but there is no effective method in the prior art to monitor the porcine erysipelas antigen in the vaccine well

Method used

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  • Selective culture medium for detecting antigen bacterium content of swine fever/swine erysipelas/swine plague triple live vaccine, and preparation method and application thereof
  • Selective culture medium for detecting antigen bacterium content of swine fever/swine erysipelas/swine plague triple live vaccine, and preparation method and application thereof
  • Selective culture medium for detecting antigen bacterium content of swine fever/swine erysipelas/swine plague triple live vaccine, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1 Accuracy check

[0032] (1) Preparation of agar plates:

[0033] Ordinary agar plate preparation: Weigh 30g TSA powder and dissolve it in 1000mL sterilized double distilled water, pressurize at 121°C for 16min, wait until it cools to 50°C, add 5% bovine serum;

[0034] Preparation of selective agar plate: Weigh 30g TSA powder and dissolve it in 1000mL sterilized double distilled water, pressurize at 121°C for 16min, wait until it cools to 50°C, add 5% bovine serum and 30mg kanamycin.

[0035] (2) Dilute erysipelas G4T10 monovalent live vaccine and porcine pneumonia EO630 monovalent live vaccine with sterile normal saline to 1 mL / head respectively, and take 100 μL and dilute them in 900 μL sterile normal saline respectively, and carry out 6 consecutive times respectively , 100 μL of the sixth dilution was inoculated on a common agar plate without kanamycin, cultured at 37°C for 36 hours, and the colonies were counted.

[0036](3) Mix erysipelas G4T10 monov...

Embodiment 2

[0042] Embodiment 2 repeatability test

[0043] (1) Prepare ordinary agar plates and selective agar plates according to the step (1) of Example 1, dilute the swine fever, swine erysipelas, porcine pneumonia triple vaccine to 1 mL / head, and take 100 μL and dilute it in 900 μL sterile normal saline , carried out 6 times in a row, and respectively took 100 μL of the sixth dilution and inoculated them on ordinary agar plates without kanamycin and selective agar plates containing kanamycin, cultured at 37°C for 36 hours, and normal Count the large colonies and small colonies in the plate respectively, and count all the colonies on the selective agar. The number of large colonies in the ordinary plate is the content of porcine pneumonia antigen, and the number of colonies in the selective plate is the content of porcine erysipelas antigen. Perform 2 repetitions as above.

[0044] (2) compare the results of two repetitions, repeat one: the number of large colonies in the common agar...

Embodiment 3

[0048] Example 3 Detection of different batches of swine fever, swine erysipelas, porcine pneumonia triple live vaccine

[0049] Test group: three different batches of swine fever, swine erysipelas, and porcine pneumonitis triple live vaccine were diluted respectively according to the method of step (1) in the embodiment of the present invention 2, inoculated, cultivated, and then counted, and the number of large colonies in the common plate was As the porcine pneumonia antigen content, the number of colonies in the selective plate is the porcine erysipelas antigen content.

[0050] Control group: Prepare ordinary agar plates and selective agar plates according to the step (1) of Example 1, dilute the swine fever, swine erysipelas, porcine pneumonia triple vaccine to 1 mL / head, and take 100 μL and dilute it in 900 μL sterile saline , carried out 6 times in a row, and respectively took 100 μL of the sixth dilution and inoculated them on ordinary agar plates without kanamycin an...

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Abstract

The invention belongs to the fields of microbiology and production of animal medicine, and particularly discloses a selective culture medium for detecting the antigen bacterium content of a swine fever / swine erysipelas / swine plague triple live vaccine, and a preparation method and application thereof. The selective culture medium comprises 29-40mg / ml of TSA (tryptone soybean agar), 3-5% of bovine serum, 20-80mu g / ml of kanamycin and a right amount of sterile double distilled water. The selective culture medium provided by the invention is simple in components and structure, convenient to prepare and easy to operate; and a determination method using the selective culture medium provided by the invention can favorably distinguish a swine erysipelas G4T10 strain and a swine plague EO630 strain in a triple vaccine, can accurately determine the swine erysipelas bacterium content, has favorable repetitiveness and high accuracy, provides a basis for monitoring stable vaccine quality during production, and is also beneficial to vaccine quality monitoring during purchase, thereby ensuring a satisfactory immune effect.

Description

technical field [0001] The invention belongs to the field of microbial production of veterinary medicine, and specifically discloses a selective culture medium for detecting the content of antigenic bacteria in a triple live vaccine of swine fever, swine erysipelas, and pneumonia, as well as a preparation method and application thereof. Background technique [0002] Swine erysipelas (SE) is an acute febrile infectious disease caused by Erysipelothrix rhusiopathiae (ER), which is characterized by high fever, acute sepsis, skin rash (subacute), chronic warty heart Meningitis and skin necrosis and polyarthritis (Wood R.L., Erysipelas, in: Leman, et al. (Eds.), Diseases of swine, Iowa State University Press, Ames, Iowa, 1992, pp.475–486 ). Most of the Erysipelothrix rhusiopathiae isolated from pigs with these symptoms were serotype 1 (subtype 1a and subtype 1b) and type 2 (Takashi et al., Protection Effect of NaOH-Extracted Erysipelothrix rhusiopathiae J.Vet.Med.Sci. 60(1):9-1...

Claims

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Application Information

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IPC IPC(8): C12Q1/06C12Q1/04C12R1/01
CPCC12Q1/045C12Q1/06
Inventor 王雷宋延华潘永飞
Owner WENS FOOD GRP CO LTD
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