Cell strain for producing goatpox vaccine

A goat pox and cell line technology, applied in the direction of microorganism-based methods, animal cells, vertebrate cells, etc., can solve the problems of lamb infection, high cost of lamb, limited number of cells, etc., and achieve the effect of low cost

Inactive Publication Date: 2015-06-10
SHANDONG LVDU BIO SICIENCE & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because the preparation of cells requires a large amount of fresh tissue, and the aseptic extraction of lamb tissue is complicated, the number of cells in the tissue is limited, and the cost of lamb is high, which severely limits the batch production of goat pox vaccine.
In order to expand the range of production cells and reduce costs, some people have tried to use primary sheep testicular cells, primary sheep kidney cells, and primary calf testicular cells to produce goat pox vaccine, but all have certain limitations.
Moreover, the current level of sheep breeding in China is different, and lambs are more likely to be infected with pathogenic bacteria and viruses such as Brucella, mycoplasma, and bovine viral diarrhea virus, which also greatly increases the biological safety risks of operators and production. inspection cost

Method used

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  • Cell strain for producing goatpox vaccine
  • Cell strain for producing goatpox vaccine
  • Cell strain for producing goatpox vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044]Comparison of Proliferation Ability of Goat Pox Virus AV41 Strain in Different Cells

[0045] (1) Virus strains and cells: goat pox virus chicken embryo attenuated strain AV41 (unpurified) was purchased from China Veterinary Drug Administration, hamster kidney cells (BHK21), pig testicular cells (ST), bovine embryonic kidney cells (MDBK) 3 kinds Cells were purchased from China Veterinary Drug Administration, and VeroB10 was screened and preserved in our laboratory. The VeroB10 cell clone is preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee, and the preservation number is CGMCC No.9705.

[0046] (2) Cell growth medium: (volume percentage) 92% DMEM, 8% newborn bovine serum, pH 7.0; cell maintenance medium: (volume percentage) 98% DMEM, 2% newborn bovine serum, The pH is 7.3.

[0047] (3) Virus proliferation: Prepare the above four kinds of cell monolayers according to the conventional method, inoculate goat pox ...

Embodiment 2

[0052] Preparation of Goat Pox Live Vaccine Using Two Passage Cells

[0053] (1) Virus strains and cells: The goat pox virus AV41 strain propagated for 1 generation on BHK21 and VeroB10 respectively was used as seed virus, and BHK21 and VeroB10 cells were used as production cells.

[0054] (2) Cell subculture and observation: After BHK21 and VeroB10 cells were digested and dispersed with 0.25% trypsin digestion solution, BHK21 cells were subcultured at a ratio of 1:4, and VeroB10 cells were subcultured at a ratio of 1:3. Cell growth medium was added and continued at 37°C. Culture until the cells grow into a monolayer.

[0055] (3) Virus inoculation and harvesting: Take BHK21 and VeroB10 cells that have formed a good monolayer, discard the cell growth medium, inoculate goat pox virus AV41 strain, add maintenance solution after adsorption for 1 hour, continue to culture at 37 °C, and wait for 75 Harvest the virus when the % cell is pathological, and the prepared cytovenom is a ...

Embodiment 3

[0060] Application Evaluation of Goat Pox Live Vaccine Prepared by Different Production Methods

[0061] (1) Experimental vaccine: goat pox vaccine prepared with two passaged cells, batch numbers VB10201301 and BHK201301 respectively; goat pox vaccine prepared with primary lamb testicular cells, batch number STC201301, all of which have passed sterility testing and specificity The inspection, exogenous virus inspection and mycoplasma inspection are qualified, and the virus content in each vaccine is not less than 10 3.5 TCID 50 .

[0062] (2) Test animals: 18 healthy goats for testing.

[0063] (3) Safety inspection: Dilute the vaccine with sterilized physiological saline so that the virus content per ml is 4×10 3.5 TCID 50 , 3 goats were injected intradermally in the chest and abdomen, each injected 2 points, 0.5ml per point, and observed for 15 days. The 3 goats vaccinated by each batch of vaccines had no obvious fluctuations in body temperature within 15 days after i...

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Abstract

The invention relates to the field of vaccine preparation, and particularly relates to a cell strain for producing a goatpox vaccine. The cell strain is an African green monkey kidney cell Vero B10 and is collected at China General Microbiological Culture Collection Center with the collection number of CGMCC No. 9705. Compared with a primary cell production method, a preparation method of the cell strain has the advantages that a goatpox virus strain AV41 is cultivated by using a VeroB10 cell, good in virus adaptability and low in strong recurrence risk; a passage cell vaccine is superior to a primary cell vaccine in terms of virus yield, uniformity, pureness and the like; the cost is low, and animals are not needed; and the safety of the passage cell vaccine is superior to that of the primary cell vaccine, and the immunity efficiency of the passage cell vaccine is not lower than that of the primary cell vaccine.

Description

technical field [0001] The invention relates to the field of vaccine preparation, in particular to a cell strain for producing goat pox vaccine and a method for producing the vaccine. Background technique [0002] Goat pox (Goatpox) and sheep pox (Sheeppox) are the pox diseases of goats and sheep caused by goat pox virus and sheep pox virus of the genus Capripoxvirus respectively. They have been popular since the 1950s. However, they are still prevalent in many countries and regions in Africa, the Middle East, Asia and Europe. In recent years, with global climate change and changes in the way animals and animal products are traded, the prevalence of these two pox diseases has become more rampant. It is the most serious pox disease that harms commercial animals. [0003] In recent years, the disease has been widely prevalent in our country. The virus can infect goats and sheep of different ages. It is characterized by fever and pox spots on the body surface or internal o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12N7/00A61K39/275A61P31/20C12Q1/70C12Q1/02G01N33/569C12R1/93
Inventor 沈志强唐娜张倩苗立中
Owner SHANDONG LVDU BIO SICIENCE & TECH
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