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Goatpox and sheep pox bivalent cell attenuated vaccine as well as preparation method and application thereof

A technology of attenuated vaccine and sheep pox virus, which is applied in the fields of goat pox and sheep pox bivalent cell attenuated vaccine and its preparation, can solve the problems of high cost of vaccine production, transportation and storage, and the heat resistance of freeze-dried protective agents, reaching The formula ingredients and preparation method are simple, the effect of ensuring immune protection and convenient transportation

Active Publication Date: 2015-07-29
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

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 pox virus bivalent cellular attenuated vaccines is still blank

Method used

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  • Goatpox and sheep pox bivalent cell attenuated vaccine as well as preparation method and application thereof
  • Goatpox and sheep pox bivalent cell attenuated vaccine as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Isolation and weakening of embodiment 1 goat pox virus (Goatpox virus HN-XY 2010)

[0036] 1.1 Isolation and identification of goat pox virus (Goatpox virus HN-XY 2010)

[0037] The inventors of the present application carried out virus isolation and case replication on goat pox-positive disease materials collected from Xinyang, Henan in 2010, and finally obtained a virus strain, which was identified by virus physicochemical characteristics and specific for goat pox virus P32 gene PCR detection and sequencing proved that the virus strain was goat pox virus virus (named Gotepox Virus HN-XY10 strain, referred to as GTPV / HN / 2010 strain).

[0038] 1.2 Attenuation of goat pox virus (Goatpox virus HN-XY2010 strain) seedling strain and establishment of virus seed batches

[0039] 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...

Embodiment 2

[0047] Isolation and attenuation of embodiment 2 sheep pox virus (Sheeppox Virus GS-WW 2010 strain)

[0048] 2.1 Isolation and identification of sheep pox virus (Sheeppox Virus GS-WW 2010 strain)

[0049] The inventor of the present application isolated the virus from sheep pox-positive disease materials collected in Wuwei, Gansu in 2010, replicated the case, and finally obtained a strain of the virus. PCR detection and sequencing proved that the virus strain was a sheep pox virus virus named (Sheeppox Virus GS-WW 2010, referred to as SPPV / GS / 2010 strain).

[0050] 2.2 Attenuation of sheep pox virus (Sheeppox Virus GS-WW 2010 strain) seedling strain and establishment of virus seed batches

[0051] The SPPV / GS / 2010 strain virus was subcultured to 17 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 head, limbs, and viscera of the chicken embryo under aseptic conditions, cut th...

Embodiment 3

[0059] Example 3 Laboratory Trial Production of Goat Pox and Sheep Pox Virus Duplex Cell Attenuated Vaccine

[0060] 3.1 Preparation of goat pox and sheep pox virus attenuated antigens

[0061] Select neonatal goat testicular primary cells (LT) that grow well and cover a single layer, pour off the nutrient solution, and inoculate goatpox virus cell passage attenuated vaccine strain Goatpox Virus HN-XY2010F43 (CCTCC NO : V201503) and sheep pox virus cell passaging attenuated vaccine strain Sheeppox Virus GS-WW 2010F44 (CCTCC NO: V201504), add DMEM maintenance solution after 10 minutes of adsorption, adjust the pH to 7.0, add penicillin and streptomycin 100 units / ml, at 37°C, 5% CO 2 Cultured in a concentration incubator. Cultivate for 48 hours, harvest when the cytopathic pathology (CPE) reaches 80%, put it in a -20°C refrigerator and freeze and thaw twice, then carry out sterility testing, safety testing, mycoplasma testing, and exogenous virus testing to meet the requireme...

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Abstract

The invention provides a goatpox and sheep pox bivalent cell attenuated vaccine. The goatpox and sheep pox bivalent cell attenuated vaccine comprises effective ingredients, namely, a goatpox virus cell passage attenuation virus and a sheep pox virus cell passage attenuation virus, wherein the goatpox virus cell passage attenuation virus is a goatpox virus cell passage attenuation vaccine strain Goatpox Virus HN-XY2010F43 strain CCTCC V201503, and the sheep pox virus cell passage attenuation virus is a sheeppox virus cell passage attenuation vaccine strain Sheeppox Virus GS-WW2010F44 strain CCTCC V201504. The goatpox and orf virus bivalent cell attenuated vaccine is safe, can have the effective immune protection force and has the important practical significance in development of safe and efficient attenuated vaccines as well as prevention and control on goatpox and sheeppox through large-scale production of goatpox and sheep pox virus bivalent cell attenuated vaccines by the aid of the attenuation virus strains.

Description

technical field [0001] The invention belongs to the technical field of biological products, and in particular relates to a goat pox and sheep pox bivalent cellular attenuated vaccine and a preparation method and application thereof. Background technique [0002] Capripox virus (CPV) is the most important animal poxvirus, belonging to the genus Capripoxvirus (Capripoxvirus) in the vertebrate poxvirus subfamily (Chordopoxrinae) of the family Poxviridae (Poxviridae) in taxonomic status, including goat poxviruses. Pox virus (Goatpox virus, GTPV), sheep pox virus (Sheep pox virus, SPPV) and bovine nodular herpes 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...

Claims

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Application Information

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IPC IPC(8): A61K39/275A61K47/42A61P31/20
Inventor 刘相涛王光祥尚佑军张志东王艳华吴锦艳陈妍田宏栾志舫逯忠新
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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