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Pseudorabies inactivated vaccine and preparation method thereof

An inactivated vaccine, pseudorabies technology, applied in vaccines, veterinary vaccines, antiviral agents, etc., can solve the problems of inability to clear the virus, difficult to produce large-scale vaccines, and backward production processes, and shorten the research and development cycle and production cycle. , the effect of maintaining the stability of viral antigens and easy large-scale production

Inactive Publication Date: 2017-03-08
CHINA ANIMAL HUSBANDRY IND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, PRV is a herpes virus, which can form persistent infection in infected pigs and carry the virus for life. Conventional attenuated vaccine immunization can control the disease, but cannot clear the virus
At present, in foreign countries, the application of attenuated vaccines is relatively common, but the virulence of attenuated vaccines that have not been sufficiently attenuated may become stronger and lead to epidemics, thereby establishing latent infection and the risk of spreading the virus; use PRV gene-deleted attenuated vaccines The prevention and control of PR lays the foundation for the eradication of pseudorabies, but the operation of this vaccine production method is relatively complicated and requires skilled researchers; the traditional production method of inactivated vaccines cannot meet the requirements for the prevention and control of pseudorabies in my country in the future. Demand control is mainly manifested in the following aspects: (1) The production cycle is long, making it difficult to achieve large-scale vaccine production in a short period of time; (2) The production process is backward, labor-intensive, and production costs are high

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Inoculation and propagation of embodiment 1 pseudorabies virus (Bartha-K61 strain)

[0028] Include the following steps:

[0029] 1. After doubling the pseudorabies virus Bartha-K61 strain with virus growth solution, inoculate the BHK-21 cells that have grown into a single layer at a ratio of 1‰, and place it at 37°C, 5% CO 2 In the incubator, culture for 24-72 hours, stop the culture after 75% of the cells appear lesions, harvest after repeated freezing and thawing 3 times, centrifuge at 12000r / min for 10min, take the supernatant, and measure the TCID of the virus liquid 50 value, and stored at -20°C after aliquoting.

[0030] 2. Suspension culture and subculture of BHK-21 cell line: Count the BHK-21 cells in suspension culture, dilute to a certain concentration with cell culture medium, inoculate into a bioreactor, and store at 37°C, 5% CO 2, pH value 7.0-7.2, dissolved oxygen 50%-60%, shaker speed 120-130rpm under the conditions of cell suspension culture for 24-72...

Embodiment 2

[0032] Inoculation and propagation of embodiment 2 pseudorabies virus (Bartha-K61 strain)

[0033] Step 3: Inoculate according to the ratio of virus to total number of cells 1:100, culture in suspension at 37°C for 24 hours with constant temperature shaking, and the rest of the operations are the same as in Example 1.

[0034] TCID of the obtained virus liquid 50 value in 10 6 TCID 50 / 0.1ml-10 8 TCID 50 In the range of / 0.1ml, the inspection is qualified, and it is stored at -20°C for later use after sub-packaging.

Embodiment 3

[0035] Inoculation and propagation of embodiment 3 pseudorabies virus (Bartha-K61 strain)

[0036] Step 3: Inoculate according to the ratio of virus to total number of cells 1:500, culture in suspension at 37°C for 48 hours with constant temperature shaking, and the rest of the operations are the same as in Example 1.

[0037] TCID of the obtained virus liquid 50 value in 10 6 TCID 50 / 0.1ml-10 8 TCID 50 In the range of / 0.1ml, the inspection is qualified, and it is stored at -20°C for later use after sub-packaging.

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Abstract

The invention provides a pseudorabies inactivated vaccine and a preparation method thereof. Pseudorabies viruses are used for inoculating BHK cells; cell suspension culture is performed in a bioreactor; a virus solution is harvested; after inactivation by formaldehyde, tween-80 is added for making a water phase; then, the water phase is mixed with an oil adjuvant or a water adjuvant and is prepared into the pseudorabies inactivated vaccine through emulsification. The BHK cell line is used for producing the pseudorabies virus (Bartha-K61 strains) inactivated vaccine for the first time; the effective measures are provided for the pseudorabies prevention and control. The pseudorabies inactivated vaccine has the advantages that the BHK cell suspension culture protection mode is used; no exogenous factor pollution is realized; the scaled production is easy; the virus antigen stability can be well maintained, and the like. The vaccine research and development period and the production period can be shortened; the vaccine support emergency capability in the sudden epidemics can be realized.

Description

technical field [0001] The invention relates to the technical field of biological products, in particular to an inactivated pseudorabies vaccine and a preparation method thereof. Background technique [0002] Pseudorabies (PR) is an acute infectious disease caused by pseudorabies virus (PRV), which is listed as a Class B infectious disease by the International Organization for Animal Health (OIE). Clinical symptoms include fever, severe itching, redness, and bleeding; paroxysmal convulsions all over the body; teeth grinding, screaming; paralysis of limbs, falling to the ground, and encephalomyelitis. Its host range is very wide. Pigs are the natural host and source of infection of the pathogen. The disease is extremely harmful to the pig industry. Tooth decay causes abortion, stillbirth, and mummified fetuses in pregnant sows, high mortality in suckling piglets, and infertility in breeding pigs. The disease was first discovered in the United States and is currently distribu...

Claims

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Application Information

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IPC IPC(8): A61K39/245A61K39/39A61K47/26A61P31/22
CPCA61K39/12A61K39/39A61K47/26A61K2039/5252A61K2039/552A61K2039/55511
Inventor 杨君敬王贺民习硕杨小蓉严石
Owner CHINA ANIMAL HUSBANDRY IND
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