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Preparation method of gosling plague inactivated vaccines as well as prepared inactivated vaccines

An inactivated vaccine and gosling plague technology, applied in the field of bioengineering, can solve the problems of infection, epidemic prevention failure, foreign virus contamination, etc., and achieve the effect of reducing the content of impurities, increasing the content of viruses, and increasing the loss of antigens

Active Publication Date: 2017-01-11
扬州优邦生物药品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Domestic commercial gosling plague vaccines are all attenuated live vaccines (SYG61 strain or GD strain), and the immunity of gosling plague live vaccines may have the following disadvantages: (1) Since GPV can only form fibroblasts in goose embryos, muscovy duck embryos, and goose Cells and muscovy duck fibroblasts are not susceptible to chicken embryos, and the goose embryos used for the production of gosling plague live vaccines are not SPF grades. There is a possibility of exogenous virus contamination in live vaccines. After immunization, There is a great risk of exogenous virus infection to geese
(2) After the geese are immunized with the goose live vaccine, the detoxification of the body will bring secondary pollution to the goose farm and hatchery, which is not good for the virus purification of the goose farm
(3) After goslings are immunized with live vaccines, it takes about one week for the immunization period to occur, and within one week goslings are the high-incidence age group for GPV infection. If the post-isolation measures are not in place, goslings are easily infected with GPV before the antibody is produced, resulting in the failure of epidemic prevention
The disadvantages of injecting yolk antibody are: (1) Each gosling is immunized after hatching, which is costly and time-consuming
(2) Since the injected yolk antibody metabolizes quickly in goslings, the duration of passive immunity is relatively short, and it can only protect goslings in the high-incidence age group, but cannot reach the whole age group of gosling plague infection in goslings. full protection
At present, there is no commercial inactivated vaccine product for the prevention and control of goose blast in my country

Method used

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  • Preparation method of gosling plague inactivated vaccines as well as prepared inactivated vaccines
  • Preparation method of gosling plague inactivated vaccines as well as prepared inactivated vaccines
  • Preparation method of gosling plague inactivated vaccines as well as prepared inactivated vaccines

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Screening of healthy susceptible goose embryos

[0025] In view of the limitations of the propagation conditions of gosling plague virus, only non-immune healthy susceptible goose embryos can be selected as the breeding carrier for seed virus preparation, seed virus production and venom production. The screening and detection of non-immune goose embryos has become one of the important tasks in the development and production of vaccines. In this study, the detection of GPV agar antibody in the yolk of non-immune goose embryos and the detection of gosling plague virus in goose embryos before inoculation ensured that non-immune goose embryos The quality of immune goose embryos, the main steps include gosling plague agar diffusion test antigen production and quality standards, detection of gosling plague agar diffusion antibody and gosling plague virus antigen detection method, and at the same time according to the appendix of the 2010 edition of "Chinese Veterinary Pharmaco...

Embodiment 2

[0067] Virus inoculation and culture preparation of virus liquid

[0068] Virus breeding

[0069] The seed poison was diluted 1:100 times with sterilized normal saline, and 11-day-old susceptible goose embryos were inoculated into the allantoic cavity, 0.2ml per embryo, and the embryos were irradiated 3-4 times a day after inoculation, and the embryos were selected to die within 48-120 hours For the goose embryos with obvious lesions, the allantoic fluid was harvested separately and placed in a sterilized container. Quantitatively subpackage the samples that have passed the sterility test and are negative for agglutination of 1% chicken erythrocyte suspension, indicate the harvest date, generation of seed virus and other information, and store them in a freezer.

[0070] inoculation

[0071] Dilute the virus seeds used for production 1:100 times with sterile saline, inoculate 11-day-old susceptible goose embryos into the allantoic cavity, 0.2ml per embryo, seal the holes wit...

Embodiment 3

[0077] Purification, concentration and vaccine preparation of virus liquid

[0078] 1 Purification of virus liquid

[0079] The qualified virus liquid is sent to the Alta Laval online disc centrifuge through a sterile pipeline, and after continuous online centrifugation, the purified virus liquid is obtained.

[0080] 2 Concentration of virus liquid

[0081] After the virus is purified, it is sent to the PALL concentrator through a sterile pipeline for concentration, and the concentration is carried out according to the concentration factor determined by the virus content before concentration, so that the virus content in each ml of the concentrated virus liquid is not less than 10 8.0 ELD 50 .

[0082] 3 Virus liquid inactivation and preparation into vaccine products

[0083] 3.1 Inactivation of virus liquid:

[0084] Filter the qualified antigen mixture with 4 layers of sterilized gauze and 1 layer of 80-mesh copper mesh, put it in an inactivation tank, add BEI cyclized...

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Abstract

The invention relates to a production method of gosling plague inactivated vaccines, and belongs to the technical field of biological products. The method comprises the following steps: (1) screening healthy and susceptible goose embryos; (2) preparing virus liquid for production; (3) determining titer of virus liquid, inactivating virus liquid, preparing vaccines, and carrying out split charging. The breeder geese are immunized, so that specific antibodies of gosling plague virus in serum and hatching egg yolks of breeder geese during egg producing period are at a high level, and after hatched goslings are hatched, the yolks are absorbed in order to produce good passive immunity protection effects. The vaccines have the advantages of high safety without hazardness due to distribution of virus, multitime immunization, substantially reduced human cost, and the like.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a preparation method and a product of gosling plague vaccine. Background technique [0002] Gosling plague virus (GPV), also known as goose parvovirus or Derzsy's pathogen, can cause acute, contact, and fatal outbreaks of goslings within 1 month of age (mainly within 20 days of age). disease. GPV belongs to the Parvoviridae family and is a member of the genus Parvovirus. It has only one serotype, but it shares some of the same antigens with Muscovy duck parvovirus (MDPV). The VP3 gene encodes the main structural protein of GPV, which can stimulate the body to produce neutralizing antibodies. Chinese scholars Fang Dingyi and Wang Yongkun first discovered the virus in Yangzhou, Jiangsu in 1956, and named it "Little Goose Plague". The Hungarian scholar Derzsy's first isolated the virus from goose embryos in 1966. In 1974, the World Poultry Association officially named it Der...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/23A61P31/20C12N7/00C12R1/93
CPCY02A50/30
Inventor 钱钟范娟宋庆庆黄海琼傅元华
Owner 扬州优邦生物药品有限公司
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