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Goatpox, sheeppox and contagious ecthyma triple cell attenuated vaccine and preparation method and application thereof

A technology of attenuated vaccine and sheep pox virus, which is applied in the field of veterinary biological products, can solve the problems of high cost of vaccine production, transportation and storage, availability of vaccines without vaccines, complex production process, etc., to ensure immune protection and simplify production process , the effect of maintaining stable quality

Active Publication Date: 2016-11-09
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
The study of goat pox, sheep and sheep aphthous triple cell attenuated vaccine is still blank

Method used

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  • Goatpox, sheeppox and contagious ecthyma triple cell attenuated vaccine and preparation method and application thereof
  • Goatpox, sheeppox and contagious ecthyma triple cell attenuated vaccine and preparation method and application thereof
  • Goatpox, sheeppox and contagious ecthyma triple cell attenuated vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Isolation, cultivation and identification of Sheep Sertoli cells

[0036] 1.1 Preparation of primary sheep testis cells

[0037] Select healthy male lambs born to healthy ewes (whose physique is healthy and negative for foot-and-mouth disease, brucellosis, and tuberculosis), and collect the scrotum after slaughter (the root of the scrotum is ligated, and the outside of the scrotum is sterilized with 75% alcohol), in an incubator, for 2 hours sent back to the laboratory. Use ophthalmic tweezers and scissors to cut the testicular parenchyma into small pieces of about 1mm×3mm, gently pipette several times in D-hanks solution, remove the supernatant, put the small tissue pieces into a 50mL centrifuge tube and add 10 times the volume of The digestion solution containing 0.1wt% type IV collagenase (GIBCO) and 0.25wt% trypsin (GIBCO) was digested at 4°C for 12h or overnight, and cultured with an equal volume of DMEM containing 10% (v / v) newborn bovine serum (GIBCO) ...

Embodiment 2

[0053] The propagation of embodiment 2 sheeppox virus

[0054] Goatpox virus can grow on tissue cells derived from cattle, sheep, and goats, and it is most sensitive to primary or secondary lamb testicular cells (LT) and lamb kidney cells (LK); after inoculation of cells with disease materials, it can grow as early as 3 days after infection A small amount of cytopathic effect (CPE) appeared, and after 4-6 days, the CPE rapidly expanded to the entire cell layer, and it was observed whether CPE appeared and lasted up to 14 days. Typical CPE is cell rounding, cell aggregation in a fascicular structure, cell shrinkage or swelling, nuclei vacuolation, chromatin fragmentation, loss of coherence in the cell monolayer, and inclusion bodies in the cytoplasm (Adla et al., 1971; Joshi et al., 1994). In addition, sheeppox virus can also propagate on BHK-21 cells and Vero cells. Some strains also grow in fetal bovine muscle cells, calf kidney cells, and bovine kidney epithelial (MDBK) ce...

Embodiment 3

[0069] Example 3 Laboratory Trial Production of Goat Pox, Sheep Pox, and Mouth Triple Cell Attenuated Vaccine

[0070] 3.1 Preparation of attenuated antigens of goat pox, sheep pox and sheep infectious impetigo virus

[0071] Select the neonatal bovine testicular Sertoli cell line NBSC (CCTCC NO: C201438, connected to BT cells) that grows well and covers a single layer, pour off the nutrient solution, and inoculate sheep infectious impetigo virus cells at 5% of the original nutrient solution volume to attenuate the virus , select natural passage cell line (LSC) of lamb testis Sertoli cells (CCTCC NO: C2016120) that grows well and covers a single layer, pour out the nutrient solution, and inoculate goat pox virus cells to subculture to attenuate Goatpox according to 5% of the original nutrient solution volume Virus HN-XY2010F43 (CCTCC NO: V201503, connected to LT cells), sheep pox virus cells passaged to attenuate Sheeppox VirusGS-WW 2010F44 (CCTCC NO: V201504, connected to LT ...

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Abstract

The invention discloses a goatpox, sheeppox and contagious ecthyma triple cell attenuated vaccine and a preparation method and application thereof. The effective components of the vaccine are separated by an inventor automatically, and after attenuation culture is conducted, goatpox virus, sheeppox virus and Orf virus cell passage attenuated viruses or passage viruses thereof are obtained, wherein a goatpox virus is a Goatpox Virus HN-XY2010F43 strain, a sheeppox virus is a Sheeppox Virus GS-WW 2010 F44 strain, and an Orf virus is an Orf Virus HB-TS09F65 strain. It is shown through attacking protection test results that the prepared goatpox, sheeppox and contagious ecthyma triple cell attenuated vaccine is safe and capable of producing an effective immune protection force, and important practical significance in developing safe and efficient goatpox, sheeppox and contagious ecthyma triple cell attenuated vaccine and preventing and controlling goatpoxes and ecthyma is achieved.

Description

technical field [0001] The invention relates to an attenuated vaccine and its preparation method and use, in particular to a goat pox, sheep pox and aphthous triple cell attenuated vaccine and its preparation method and use. The invention belongs to the field of veterinary biological products. 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 Pox virus (Goatpox virus, GTPV), sheep pox virus (Sheep poxvirus, 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 rat...

Claims

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Application Information

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IPC IPC(8): A61K39/295A61P31/20
CPCA61K39/12A61K2039/70
Inventor 尚佑军张志东王光祥刘湘涛王艳华吴锦艳陈妍张克山田宏尹双辉
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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