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Goatpox virus-orf virus combined cell attenuated vaccine and its preparation method and use

An attenuated vaccine and aphtha virus technology, which are applied in the field of goat pox, attenuated vaccine and preparation thereof, and aphthous double cell attenuated vaccine and preparation thereof, and can solve the problem of vaccines with no vaccine, complicated production process, vaccine production and transportation. And the problem of high storage cost, to achieve the effect of convenient transportation, broad application prospects, and guaranteeing immune protection.

Active Publication Date: 2015-07-08
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In China, although the attenuated oral ulcer vaccine has been developed in the Veterinary Research Institute of the Qinghai Academy of Animal Husbandry and Veterinary Sciences and the Gansu Provincial Animal Husbandry and Veterinary Research Institute, the production process is complex and backward, and the freeze-dried protective agent is not heat-resistant and many other production process bottlenecks. It is because the vaccine cannot be mass-produced, and the vaccine is currently in a situation where there is no vaccine available
In addition, the attenuated virus is a strain from around the 1980s, and the current domestic epidemic virus may mutate, making it difficult to guarantee the effectiveness of its immunity
Goat pox attenuated vaccines are produced by several domestic vaccine manufacturers, but the production process of the vaccine requires primary cells of sheep testis, and technical problems such as freeze-drying protective agents are not heat-resistant, which make the production, transportation and storage costs of the vaccine too high
Research on goat pox and sheep aphthous virus dual cell attenuated vaccine is still blank

Method used

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  • Goatpox virus-orf virus combined cell attenuated vaccine and its preparation method and use
  • Goatpox virus-orf virus combined cell attenuated vaccine and its preparation method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Isolation and identification of sheep infectious pustular virus (Orf Virus HB-TS09 strain), attenuation of seed virus, establishment of seed batches and development of heat-resistant freeze-drying protective agent

[0025] Isolation, identification, attenuation, establishment of seed batches and development of a heat-resistant freeze-drying protective agent for sheep infectious pustules virus strain (Orf Virus HB-TS09 strain) were described in the inventor's invention patent application: "A Sheep Infectious Disease" Infectious impetigo virus cell attenuated vaccine and its preparation method and application" (Application No.: CN201410160855.0, Publication No.: CN104017776A) and invention patent application "Heat-resistant freeze-drying protectant for goat infectious impetigo virus cell attenuated vaccine and its preparation Methods and Applications" (Application No.: CN201410101828.6, Publication No.: CN103893776A) have been described. The inventor has success...

Embodiment 2

[0029] Isolation and weakening of embodiment 2 goat pox virus (Goatpox virus HN-XY 2010)

[0030] 2.1 Isolation and identification of goat pox virus (Goatpox virus HN-XY 2010)

[0031] The inventor of the present invention collected goat pox-positive disease materials from Xinyang, Henan in 2010, isolated the virus, replicated the case, and finally obtained a virus strain, which was identified by virus physicochemical characteristics and specific PCR for the P32 gene of goat pox virus. Detection and sequencing proved that the virus strain is goat pox virus virus (named Gotepox Virus HN-XY10 strain, referred to as GTPV / HN / 2010 strain).

[0032] 2.2 Attenuation of goat pox virus (Goatpox virus HN-XY2010 strain) seedling strain and establishment of virus seed batches

[0033] The GTPV / HN / 2010 strain virus was subcultured to 16 passages with the primary cells of sheep testis, and the GTPV virulence lgTCID 50 ≥3.5, choose 9-day-old to 11-day-old SPF chicken embryos, remove the he...

Embodiment 3

[0041] Example 3 Laboratory Trial-manufacture of Goat Pox and Mouth Virus Dual Cell Attenuated Vaccine

[0042] 3.1 Preparation of attenuated antigens of goat pox and sheep infectious impetigo virus

[0043] Select primary cells of newborn sheep testis (LT) and primary cells of newborn bovine testis (BT) that have grown well and cover a single layer, pour off the nutrient solution, and inoculate goat pox virus cells to subculture to attenuate the virus according to 5% of the original nutrient solution volume Goatpox Virus HN-XY2010F43 (CCTCC NO: V201503) and Orf Virus HB-TS09F65 virus liquid (CCTCC NO: V201406), which were passaged by sheep infectious impetigo virus cells, were absorbed for 10 minutes and then added with DMEM maintenance solution to adjust the pH to 7.0 , add 100 units / ml each of penicillin and streptomycin, and cultivate in a 37°C, 5% CO2 concentration incubator. Cultivate for 48 hours, harvest when the cytopathic pathology (CPE) reaches 80%, put it in a -20...

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Abstract

The invention discloses a goatpox virus-orf virus combined cell attenuated vaccine and its preparation method and use. The goatpox virus-orf virus combined cell attenuated vaccine contains an active component which comprises goatpox and contagious pustular dermatitis virus cell passage-attenuated viruses or passaged viruses separated by the inventor and subjected to passage attenuation culture, wherein the goatpox virus cell passage-attenuated virus is a Goatpox Virus HN-XY2010F43 strain and has a preservation number of CCTCC NO: V201503 and the contagious pustular dermatitis virus cell passage-attenuated virus is an Orf Virus HB-TS09F65 strain and has a preservation number of CCTCC NO: V201406. A toxin counteracting protection test result shows that the goatpox virus-orf virus combined cell attenuated vaccine is safe and can produce an effective immunization protection function. The goatpox virus-orf virus combined cell attenuated vaccine and its preparation method have important practical meanings for development of safe and efficient attenuated vaccines and large-scale production of the goatpox virus-orf virus combined cell attenuated vaccine for preventing and controlling goatpox and orf from the disclosed attenuated strain.

Description

technical field [0001] The invention relates to an attenuated vaccine and its preparation method and use, in particular to a goat pox and aphthous double cell attenuated vaccine and its preparation method and use. The invention belongs to the field of veterinary biological products. Background technique [0002] Capripoxvirus (CPV) is a member of the Capripoxvirus genus of the Poxviridae Vertebratepoxvirinae subfamily. Members of this genus include goat pox virus (GTPV), sheep pox virus (SPV) and bovine nodular rash virus (Lumpy skin disease virus, LSDV). Goatpox is an acute, contagious infectious disease caused by goatpox virus. It is listed as one of the 93 animal diseases that must be reported by the World Organization for Animal Health (FAO / OIE), and it is listed as a first-class infectious disease in my country. Its morbidity rate can reach more than 75%, the mortality rate can reach more than 50%, and the lamb mortality rate can reach 100%, and it can also cause abort...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/275A61K47/42A61P31/20
Inventor 刘湘涛王光祥尚佑军张志东王艳华田红吴锦艳陈妍栾志舫逯忠新
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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