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Porcine pseudorabies virus strain as well as inactivated vaccine and applications thereof

A technology of porcine pseudorabies virus and porcine pseudorabies, applied in antiviral agents, viruses/bacteriophages, biochemical equipment and methods, etc., can solve the problems of poor immune induction effect, backward inactivation process, residual inactivation of inactivation agents, etc. problems, to achieve good immune protection efficacy and safety, good cell proliferation activity, and good immunogenicity

Active Publication Date: 2013-09-18
泰州博莱得利生物科技有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The existing porcine pseudorabies virus inactivated vaccine mainly has the following problems: the one is that the virus strain is aging and the virus titer is not high; Is the use of traditional adjuvants, resulting in poor immune induction effect

Method used

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  • Porcine pseudorabies virus strain as well as inactivated vaccine and applications thereof
  • Porcine pseudorabies virus strain as well as inactivated vaccine and applications thereof
  • Porcine pseudorabies virus strain as well as inactivated vaccine and applications thereof

Examples

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Effect test

Embodiment 1

[0033] The implementation of the present invention will be described in more detail through the following examples, but it should be understood that the examples are only exemplary and do not constitute any limitation to the scope of the present invention. Those skilled in the art should understand that the details and forms of the technical solution of the present invention can be modified or replaced without departing from the spirit and scope of the present invention, but these modifications or replacements all fall within the protection scope of the present invention. Isolation, cultivation and identification of porcine pseudorabies virus BJ strain of embodiment 1

[0034] 1. Isolation and culture characteristics of porcine pseudorabies virus BJ strain

[0035] (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 ...

Embodiment 2

[0047] The preparation of embodiment 2 porcine pseudorabies virus BJ strain seed virus

[0048] 1. Establishment of basic seed batches: Take the isolated and identified original virus seeds, insert them into cell cultures such as ST cells, PK15 cells, MDCK cells, or BHK-21 cells that form a single layer according to 1% of the amount of virus culture solution, and place 5%CO 2 Culture at 37°C, and observe CPE every day. Observation results show that: the cells swell, shrink, and then begin to fall off and gradually form plaque lesions, and there is a phenomenon of "pulling the net". When the lesion reaches 80%, the toxic cell culture medium is harvested, and after 2 times of freezing and thawing, the toxin is collected . Continuous passage for 6 generations.

[0049] 2. Establishment of production seed batches: take basic seeds, insert 1% of the amount of virus culture solution into cell cultures such as ST cells, PK15 cells, MDCK cells or BHK-21 cells that form a single lay...

Embodiment 3

[0061] The preparation of embodiment 3 porcine pseudorabies virus disease inactivated vaccines

[0062] 1. Preparation of virus liquid for seedling preparation: the porcine pseudorabies virus BJ strain virus seed separated by embodiment 1 is cultivated in the manner of embodiment 2 to produce seed poison, and 0.1% of the virus culture liquid amount is inserted to form a monolayer In cell cultures such as ST cells, PK15 cells, MDCK cells or BHK-21 cells, put them in 37°C rotary culture. When the lesion reaches 80%, harvest the toxic cell culture medium, and collect the poison after 3 times of freezing and thawing.

[0063] 2. Inactivation: Add 2% (w / v) divinylimine solution to the virus liquid according to 0.05% (w / v) of the total amount of virus liquid, under the condition of 32°C and 120r / min air bath shaking , inactivated for 48h, then add 2% sodium thiosulfate solution to terminate the inactivation, and do a sterility test.

[0064] 3. Concentration: get the inactivated vi...

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Abstract

The invention discloses a porcine pseudorabies virus strain as well as an inactivated vaccine and applications thereof, belonging to the field of separation and application of the porcine pseudorabies virus strain. The invention firstly provides a porcine pseudorabies virus BJ strain separated from diseased pig tissues, and the microbial preservation number of the porcine pseudorabies virus BJ strain is CGMCC (China General Microbiological Culture Collection Center) No.7351. The invention discloses a method for preparing the inactivated vaccine by applying the porcine pseudorabies virus BJ strain. The method comprises the steps of culturing a virus strain to obtain a virus solution; adding an inactivator, and inactivating and concentrating the virus solution; and evenly mixing an adjuvant and the virus solution, and emulsifying to obtain the inactivated vaccine. The technological parameters of the inactivated vaccine preparation method are further optimized, and the immune protection efficacy and safety of the inactivated vaccine can be improved. Shown by the immune protection efficacy and safety tests, the porcine pseudorabies inactivated vaccine prepared has good immune protection efficacy and safety, and can be clinically used for preventing or treating porcine pseudorabies.

Description

technical field [0001] The present invention relates to an isolated virus epidemic strain, in particular to an isolated porcine pseudorabies epidemic virus strain and an inactivated porcine pseudorabies vaccine prepared from the strain, belonging to the isolation and purification of porcine pseudorabies virus strains. application field. Background technique [0002] Pseudorabies virus (Porcine pseudorabies virus, PRV) can cause a variety of diseases in domestic and wild animals with fever, itching (except pigs) and encephalomyelitis as the main symptoms. Immunization is the main strategy for preventing and controlling pseudorabies. Currently, the Bartha-K61 strain is widely used in China to prepare attenuated vaccines. Although the attenuated PRV gene deletion vaccine can effectively prevent the emergence of clinical symptoms after infection, the vaccinated pigs may still be infected by the strong virus, and the infection can form a latent virus, and then the latent infecte...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/245A61P31/22C12R1/93
Inventor 刘继红
Owner 泰州博莱得利生物科技有限公司
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