A kind of vaccine composition and its preparation method and application

A vaccine composition and composition technology, applied in the field of vaccine composition, can solve the problems that antibacterial drugs are difficult to work, vaccines are difficult to achieve immune effect, and pig immunity is reduced

Active Publication Date: 2016-09-14
PULIKE BIOLOGICAL ENG INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the only vaccines against the infection of these three pathogens are porcine reproductive and respiratory syndrome, mycoplasma hyopneumoniae and Haemophilus parasuis single vaccines, and it is impossible to achieve a vaccine to resist the mixed infection caused by these three pathogens
[0006] As the pathogenic strains and mixed infections have made it difficult for existing vaccines to achieve the expected immune effect, the problem now is that if pigs are infected or inoculated with porcine reproductive and respiratory syndrome virus, the immune effect of the mycoplasma pneumoniae vaccine will be obvious Reduced, Yang Hanchun ("Epidemic characteristics and control strategies of pig immunosuppressive diseases", China Animal Husbandry and Veterinary Medicine, 2004, 31 (5): 41-43), Wan Suiru ("Main technical measures for preventing and controlling pig immunosuppression", Pig, 2009, 1: 42-46) and other studies found that porcine reproductive and respiratory syndrome is an immunosuppressive disease; Symposium Proceedings, pp. 262-265, 2008) and others found that porcine reproductive and respiratory syndrome virus infection reduces immunity in pigs (eg, by disrupting the normal function of alveolar macrophages) and induces immunosuppression
The sick pigs showing these symptoms are caused by the synergistic action of these three pathogens, and once secondary infection with Haemophilus parasuis, various antibacterial drugs are difficult to be effective

Method used

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  • A kind of vaccine composition and its preparation method and application
  • A kind of vaccine composition and its preparation method and application
  • A kind of vaccine composition and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The preparation of embodiment 1 porcine reproductive and respiratory syndrome virus live vaccine

[0056] In this example, the spinner bottle culture method was used. Marc-145 cells were expanded and cultured in spinner bottles with cell growth medium (MEM medium containing 5% bovine serum, pH 7.25). The cells were cultured until the fifth day, and the PRRS virus was inoculated according to the M.O.I. of 0.001. After culturing until the third day after inoculation, the virus liquid was harvested and stored at -20°C. According to the highly pathogenic porcine reproductive and respiratory syndrome live vaccine (JXA1-R strain) testing procedures. Determination of virus content was 10 8.0 TCID 50 / ml.

[0057] Mix the qualified virus liquid in the same container, add 2% gelatin, 5% dextrin, 10% trehalose, 2% polyvinylpyrrolidone (PVP), 2% bovine serum at a ratio of 1:1 (V / V) Albumin, 0.164% Potassium Dihydrogen Phosphate, 0.052% Disodium Hydrogen Phosphate and water a...

Embodiment 2

[0059] Embodiment 2 Haemophilus parasuis disease, Mycoplasma hyopneumonia inactivated vaccine preparation

[0060] Haemophilus parasuis disease and Mycoplasma hyopneumoniae inactivated vaccine, composed of inactivated, concentrated and security-checked Haemophilus parasuis serotype 4 JS strain, type 5 ZJ strain antigen and Mycoplasma hyopneumoniae HN0613 strain antigen, and Montanide GEL The ST adjuvant is mixed and prepared, the mixed antigen accounts for 75% to 90% of the total volume of the vaccine, and the adjuvant content is 10% to 25%.

[0061] 1. Preparation of Haemophilus parasuis and Mycoplasma hyopneumoniae seeds

[0062] 1.1 Preparation of Haemophilus parasuis seeds

[0063] Haemophilus parasuis serotype 4 JS strain and type 5 ZJ strain freeze-dried strains were streaked and inoculated on the preferred Haemophilus parasuis medium (MHPs) plate containing agar powder in our laboratory, and placed at 37°C Cultivate for 18-24 hours, select the colonies that meet the r...

Embodiment 3

[0094] Example 3 Porcine Reproductive and Respiratory Syndrome Live Vaccine-Mycoplasma hyopneumoniae, Haemophilus parasuis Inactivated Vaccine Correlation Test

[0095] 1. Physical property test, sterility test

[0096] One batch of porcine reproductive and respiratory syndrome live vaccine - Mycoplasma hyopneumoniae and Haemophilus parasuis inactivated vaccine (batch number 120701), passed the physical property test and sterility test. The detailed results are shown in Table 2:

[0097] Table 2 Vaccine physical properties test and sterility test

[0098]

[0099] Note: T.G means thioglycolate medium, G.A means casease agar medium, G.P glucose-peptone medium; "-" means sterile growth.

[0100] 2 safety test

[0101] Use 5 piglets negative for porcine reproductive and respiratory syndrome virus, Haemophilus parasuis antigen, and Mycoplasma hyopneumoniae antibody at the age of 3 to 4 weeks, and inoculate each muscle with 10 parts of live vaccine diluted with 2 parts of inac...

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Abstract

The present invention provides a vaccine composition comprising an immune amount of Mycoplasma hyopneumoniae antigen, an immune amount of a Haemophilus parasuis antigen, and an immune amount of an antigen of Porcine Reproductive and Respiratory Syndrome Virus. The vaccine composition is infected with porcine reproductive and respiratory syndrome virus, Mycoplasma hyopneumoniae and Haemophilus parasuis on the 14th day after the second immunization, and can protect more than 80% of pigs without mutual interference between antigens, and achieves impressive results. Satisfactory immune effect.

Description

technical field [0001] The invention relates to a vaccine composition, which is used for preventing and treating porcine respiratory disease syndrome. Background technique [0002] Porcine reproductive and respiratory syndrome (Porcine Reproductive and Respiratory Syndrome, PRRS), also known as porcine blue ear disease, is caused by single-stranded positive-sense RNA porcine reproductive and respiratory syndrome virus. Stillbirths, mummies, etc. and pigs of all ages, especially piglets, are characterized by respiratory diseases, and have now become one of the main diseases in large-scale pig farms. The highly pathogenic porcine blue-ear disease that broke out in some parts of my country in 2006 is an acute and highly lethal disease caused by a mutant strain of porcine reproductive and respiratory syndrome virus, which belongs to the second category of animal diseases in the country. The disease has new epidemic characteristics in our country, the virus has mutated, the path...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/295G01N33/53A61P11/00
Inventor 张许科孙进忠白朝勇
Owner PULIKE BIOLOGICAL ENG INC
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