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Vaccine composition, kit, preparation methods of vaccine composition and kit, and applications of vaccine composition and kit

A vaccine composition and adjuvant technology, applied in the directions of antiviral agents, pharmaceutical formulations, medical preparations containing active ingredients, etc., can solve problems such as inability to treat kennel cough, achieve excellent immune effect and reduce morbidity Effect

Inactive Publication Date: 2017-08-11
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this invention is aimed at multiple diseases of dogs, and cannot carry out targeted treatment for kennel cough (two diseases)

Method used

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  • Vaccine composition, kit, preparation methods of vaccine composition and kit, and applications of vaccine composition and kit
  • Vaccine composition, kit, preparation methods of vaccine composition and kit, and applications of vaccine composition and kit
  • Vaccine composition, kit, preparation methods of vaccine composition and kit, and applications of vaccine composition and kit

Examples

Experimental program
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Effect test

Embodiment approach

[0040] As a preferred embodiment of the present invention, in the vaccine composition of the present invention, the gene sequence of the F protein includes the accession numbers disclosed in NCBI as AY491509.1, EF546392.1, EF487542.1, AX586947. 1. F protein gene sequence of KC237065.1, KC237064.1, KC237063.1;

[0041] The gene sequence of the HN protein includes, but is not limited to, the gene sequences of the HN protein with the accession numbers EF543647.1, KC237065.1, KC237064.1, KC237063.1, and EF546393.1 published in NCBI;

[0042] The gene sequence of the NP protein includes, but is not limited to, NP protein gene sequences published in NCBI with accession numbers EF543648.1, EF546391.1, KC237065.1, KC237064.1, KC237063.1, and AY581307.1.

[0043] As a most preferred embodiment of the present invention, in the vaccine composition of the present invention, the F, HN, and NP protein antigen sequences of canine parainfluenza virus are the F, HN, and NP proteins whose NCBI ...

Embodiment 1

[0073] Example 1 Preparation and Content Determination of Canine Parainfluenza Virus Antigen

[0074] 1.1 Preparation and content determination of CPIV live antigen

[0075] After diluting the CPIV Cornell strain with DMEM culture medium, inoculate a well-grown Vero cell monolayer at 5% V / V of the culture medium volume, absorb at 37°C for 30 minutes, add DMEM culture medium, and place at 37°C, 5% CO 2 Cultivate in an incubator, and when 80% of the cells have lesions, freeze and thaw twice at -20°C to harvest the virus liquid. The harvested CPIV Cornell strain virus solution was diluted serially by 10 times, and 10 -3 、10 -4 、10 -5 、10 -6 4 dilutions, each dilution was inoculated into 5 wells of a 96-well cell culture plate, 0.1ml per well, and a negative control was set up at the same time, at 37°C, 5% CO 2 Cultivate in the incubator 7 days, observe cytopathic change (CPE), calculate according to Reed-Muench method and draw CPIV virus content to be 10 7.5 TCID 50 / ml. ...

Embodiment 2

[0084] Example 2 Preparation and content determination of Bordetella canine bronchiseptica antigen

[0085] 2.1 Preparation and content determination of live Bb antigen

[0086] After rethawing the freeze-dried strain of Bb SD25d strain with physiological saline, streak inoculate it on a TSA plate containing 5% newborn bovine serum, incubate at 37°C for 24 hours, observe the colony shape, pick 5-10 typical colonies, and streak Inoculate on TSA slant containing 5% newborn bovine serum, culture at 37°C for 24 hours, store as primary seeds at 2-8°C, and use for no more than 2 weeks. Take first-grade seeds and inoculate 1% of TSB liquid medium containing 5% newborn bovine serum, 37°C, 200 rpm, shake culture for 16 hours, take samples for pure inspection, and use them as second-grade seeds after passing the test, and the use period shall not exceed 2 week. Take secondary seeds, inoculate according to 1% of the medium volume, ferment and cultivate at 37°C for 24 hours to harvest t...

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Abstract

The invention provides a vaccine composition. The vaccine composition comprises a dog parainfluenza virus antigen with the immunity effective dose, a dog bordetella bronchiseptica antigen with the immunity effective dose, and an adjuvant, wherein the dog parainfluenza virus antigen is selected from an inactivated dog parainfluenza virus antigen or at least one of F protein, HN protein, M protein, NP protein, P protein and L protein, and the bordetella bronchiseptica antigen is selected from inactivated dog bordetella bronchiseptica antigen or p68 protein. Compared with the vaccines in the prior art, the vaccine composition provided by the invention has the better immune effect, and has no side effects, and thus the risk that a veterinarian worker or a contactor is infected is reduced. The invention further relates to a kit. The kit contains the inactivated dog parainfluenza virus antigen or the subunit antigen, the attenuated live dog bordetella bronchiseptica antigen or the attenuated live dog parainfluenza virus antigen, and the inactivated dog bordetella bronchiseptica antigen or p68 protein.

Description

technical field [0001] The invention belongs to the field of veterinary medicine, and in particular relates to a vaccine composition containing canine parainfluenza virus antigen and canine bronchiseptica Bordetella antigen, a kit, a preparation method and application thereof. Background technique [0002] Canine Infectious Tracheobronchitis (ITB), also known as Kennel cough, is an acute and highly contagious respiratory infectious disease of dogs. Highly lethal pathogen infection and other characteristics, only in the survey of Gansu Province from 2004 to 2009, the overall incidence of the disease was as high as 34.26% (Lu Wangyin. Epidemiological investigation and prevention of kennel cough in Gansu Province. Journal of Animal Husbandry and Veterinary Medicine, 2010, 29(3):49-52). The disease mainly causes sudden paroxysmal coughing of puppies accompanied by variable nasal secretions, which often occur frequently in spring and autumn. Once the disease occurs in the dog gr...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61K39/145A61P31/16A61P31/04A61K39/10
Inventor 张许科孙进忠韩水仲田克恭
Owner PU LIKE BIO ENG
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