Porcine pseudorabies virus strain and porcine pseudorabies inactivated vaccine prepared by using same

A technology of porcine pseudorabies virus and porcine pseudorabies, which is applied in the direction of antiviral agents, viruses/phages, and medical preparations containing active ingredients, etc., to achieve the effects of fast proliferation, high titer, and good immunogenicity

Active Publication Date: 2012-02-08
哈药集团生物疫苗有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For porcine pseudorabies virus, the application of diethyleneimine inactivation, and the...

Method used

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  • Porcine pseudorabies virus strain and porcine pseudorabies inactivated vaccine prepared by using same
  • Porcine pseudorabies virus strain and porcine pseudorabies inactivated vaccine prepared by using same
  • Porcine pseudorabies virus strain and porcine pseudorabies inactivated vaccine prepared by using same

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Experimental program
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Embodiment 1

[0034] Isolation, cultivation and identification of porcine pseudorabies virus H strain of embodiment 1

[0035] 1. Isolation and cultivation of porcine pseudorabies virus H strain

[0036] 1. Disease material collection: Aseptically collect the brain, tonsil and other tissues of stillborn sows. Add MEM at a ratio of 1:10, grind, and prepare tissue suspension. After repeated freezing and thawing for 3 times, centrifuge at 2000r / min for 15min, collect the supernatant, and then filter through a 0.2um membrane filter, and store the filtrate at -40°C. Store in the refrigerator.

[0037] 2. Separation and culture: Inject the above-mentioned disease material into the cell culture that has not yet formed a monolayer according to the content of 10% of the virus culture solution, incubate at 37°C for 1 hour, change to MEM culture solution containing 2% calf serum, and keep at 37°C Cultivate for 5 days. The second generation was cultivated for 96 hours, and the poisonous culture solu...

Embodiment 2

[0050] The preparation of embodiment 2 porcine pseudorabies virus disease inactivated vaccines

[0051] 1. Preparation of virus liquid for seedling preparation: the porcine pseudorabies virus H strain virus seed separated by embodiment 1 is inserted into the ST cell culture forming a monolayer by 1% of the virus culture liquid amount, and put 37 ℃ of rotation cultures , when the lesion reaches 80%, harvest the toxic cell culture medium, and collect the poison after 2 times of freezing and thawing.

[0052] 2. Inactivation: Add 2% (w / v) divinylimine solution to the virus liquid according to 0.02% (w / v) of the total amount of virus liquid, shake it fully, and inactivate it on a shaker at 30°C and 100r / min. Live for 60h, then add 2% sodium thiosulfate solution to terminate the inactivation. And for sterility test.

[0053] 3. Concentration: Take the inactivated virus liquid, centrifuge it horizontally at 4000r / min, and then filter it with a 0.2um membrane filter. The virus liq...

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Abstract

The invention discloses a porcine pseudorabies virus strain and a porcine pseudorabies inactivated vaccine prepared by using the same. The porcine pseudorabies virus strain is separated from the brain, tonsil and other tissues of a still birth of a sow on a pig farm by virtue of subculture adaptation and has a microbial preservation number of GCMCC (China General Microbiological Culture Collection Center) No.5013. The virus strain separated in the invention has quick proliferation and high titer on ST (scheduled tribe) cells, can induce an animal to generate a high-titer neutralizing antibody, and has excellent immunogenicity. The invention also discloses a preparation method of the porcine pseudorabies inactivated vaccine, and the method comprises the following steps of: culturing the porcine pseudorabies virus strain to obtain a virus solution; adding an inactivator to inactivate the virus solution; and preparing an oil phase and a water phase, and emulsifying, thus obtaining the porcine pseudorabies inactivated vaccine. In the invention, each process parameter of the inactivated vaccine is optimized. Immune protective efficacy and safety tests prove that the inactivated vaccine has excellent immunogenicity and safety.

Description

technical field [0001] The invention relates to an isolated virus strain, in particular to an isolated porcine pseudorabies virus strain and an inactivated porcine pseudorabies vaccine prepared by using the strain. The invention belongs to the field of porcine pseudorabies inactivated vaccines. Background technique [0002] Porcine pseudorabies is an acute infectious disease caused by porcine pseudorabies virus (PRV), characterized by fever, itching and encephalomyelitis. Porcine pseudorabies virus commonly causes abortion, mummified fetuses, stillbirths and weak litters in pregnant sows. Newborn piglets have neurological symptoms, and the infection rate and mortality rate can reach 100%. Adult pigs are more resistant to infection after infection, showing recessive infection, resulting in long-term virus infection and detoxification, becoming the most dangerous source of infection, seriously affecting the production of breeding pigs and fine breeds promotion. With the deve...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/245A61P31/22C12R1/93
Inventor 姜力付丽杰任德强吴金戴秀莉张智明武佳斌潘兴广张立恒丁国杰李尊江张杨赵辉
Owner 哈药集团生物疫苗有限公司
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