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A kind of porcine pseudorabies virus mutant xf-1 strain and its preparation method and application

A technology for porcine pseudorabies and virus mutation, applied in biochemical equipment and methods, viruses, antiviral agents, etc., can solve problems such as differences in immunogenicity, achieve high antibody levels, better safety, and reduce labor costs.

Active Publication Date: 2019-04-02
WUHAN KEQIAN BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the continuous mutation of the epidemic strains, there are differences in immunogenicity between the commercially available vaccines and the vaccines prepared by traditional strains, which cannot prevent and control the current epidemic of porcine pseudorabies.

Method used

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  • A kind of porcine pseudorabies virus mutant xf-1 strain and its preparation method and application
  • A kind of porcine pseudorabies virus mutant xf-1 strain and its preparation method and application
  • A kind of porcine pseudorabies virus mutant xf-1 strain and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Preparation of porcine pseudorabies virus (XF-1 strain) liquid and vaccine:

[0037] 1. Suspension culture of BHK-21 cells in a bioreactor:

[0038] Take 5ml of BHK-21 seed cells preserved in liquid nitrogen, melt rapidly in a 37°C water bath, centrifuge at 1000rpm for 5 minutes, discard the supernatant, resuspend with 100ml of BHK medium containing 2% (v / v) low serum, and transfer to In a 250ml shake flask, culture on a shaker at a constant temperature at 37°C with a rotation speed of 80-100rpm / min for suspension culture for 2 days until the cell density reaches 2.0-4.0×10 6 cells / ml, according to the ratio of 1:4, at 37 ° C, 80 ~ 100 rpm for 2 ~ 3 days, the cells were expanded to a volume of 1000 ml, so that the cell density reached 4.0 ~ 6.0 × 10 6 cells / ml in order to obtain BHK-21 cell suspension.

[0039] Inoculate 1000ml of the BHK-21 cell suspension obtained above into a 10L bioreactor, add 8L of BHK medium containing 2% (v / v) newborn bovine serum, control the...

Embodiment 2

[0047] Immunogenicity Test of Porcine Pseudorabies Inactivated Vaccine (XF-1 Strain)

[0048] 1. Materials and methods

[0049] 1.1 The experimental animal market purchased 21-25-day-old piglets from a pig farm in Wuhan, and tested the main pathogens and related antibodies. A total of 30 piglets with negative neutralizing antibody against pseudorabies (PRV neutralizing antibody titer not higher than 1:2).

[0050] 1.2 The challenge strain The field strain of porcine pseudorabies virus (XF strain) comes from Wuhan Keqian Biotechnology Co., Ltd.

[0051] 1.3 Method The test pigs were randomly divided into 6 groups, 5 heads in each group, and the conditions of each group were as follows:

[0052] The 1st group is porcine pseudorabies inactivated vaccine (XF-1 strain), promptly selects the vaccine that embodiment 1 prepares;

[0053] The second group is imported live vaccine (Bartha-K61 strain), batch number: 77BN, purchased from the Spanish Haibolai Biopharmaceutical Factory; ...

Embodiment 3

[0074] Comparative experiment on the growth and decline of antibodies against several different porcine pseudorabies vaccines

[0075] 1. Test materials and methods

[0076] 1.1 The experimental animal market purchased 21-25-day-old piglets from a pig farm in Wuhan, and tested the main pathogens and related antibodies. A total of 25 piglets with negative neutralizing antibody against pseudorabies (PRV neutralizing antibody titer not higher than 1:2).

[0077] 1.2 The neutralizing titer of porcine pseudorabies-specific positive serum is 1:256, provided by Wuhan Keqian Biotechnology Co., Ltd.

[0078] 1.3 Test method The test pigs were randomly divided into 5 groups, 5 heads in each group, and the grouping conditions were as follows:

[0079] The first group selects the vaccine prepared in Example 1 for porcine pseudorabies inactivated vaccine (XF-1 strain);

[0080]The second group is the imported live vaccine (Bartha-K61 strain), batch number: 77BN, purchased from the Spani...

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Abstract

The invention relates to the technical field of swine pseudorabies viruses, and relates to a swine pseudorabies virus variant XF-1 strain, a preparing method thereof and applications of the strain. The accession number of the variant is CCTCC NO V201654. The swine pseudorabies virus variant (the XF-1 strain) is adopted to prepare an inactivated vaccine, and safety and immunoprotective experiments are performed. Results prove that the swine pseudorabies virus variant (the XF-1 strain) inactivated vaccine has good safety, and that the immunoprotection efficiency of the inactivated vaccine is obviously higher than that of other immune groups, and therefore the virus strain is proved to have good immunogenicity, can be used for vaccine research and development for the swine pseudorabies, and has a food vaccine development prospect.

Description

technical field [0001] The invention mainly relates to the technical field of porcine pseudorabies inactivated vaccines, in particular to a porcine pseudorabies virus mutant XF-1 strain, a preparation method and application thereof. Background technique [0002] Pseudorabies is an acute infectious disease caused by pseudorabies virus (Pseudorabies virus, PRV), including a variety of livestock and wild animals. Pigs are the natural host and reservoir of the virus. The disease is characterized by fever, severe itching, encephalomyelitis and severe reproductive impairment. Since 2011, many provinces across the country have used gE gene-deleted live vaccines for large-scale immunization. Weak piglets, stillbirths, mummies, and miscarriages occur in sows in pig farms; gilts and empty sows often appear to return to estrus, repeated infertility or anoestrus; boars often suffer from testicular atrophy and loss of sexual function after infection breeding ability; new piglets have ne...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/245A61P31/22C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552C12N7/00C12N2710/16721C12N2710/16734C12N2710/16751
Inventor 陈波徐高原周明光方雨林曾爱星陈章表金建云
Owner WUHAN KEQIAN BIOLOGY CO LTD
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