Preparation method and application of newcastle disease virus living-vector vaccine through gene recombination of canine distemper attenuated vaccine strains F and H

A technology of canine distemper virus and attenuated vaccine, applied in the direction of virus/bacteriophage, antiviral agent, virus antigen components, etc., can solve problems such as interference with immune effect and immune failure

Inactive Publication Date: 2012-12-12
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] CD can be prevented, mainly by inoculating puppies with canine distemper attenuated vaccine, but the existence of maternal antibodies will directly interfere with the immune effect, often leading to immune failure

Method used

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  • Preparation method and application of newcastle disease virus living-vector vaccine through gene recombination of canine distemper attenuated vaccine strains F and H
  • Preparation method and application of newcastle disease virus living-vector vaccine through gene recombination of canine distemper attenuated vaccine strains F and H
  • Preparation method and application of newcastle disease virus living-vector vaccine through gene recombination of canine distemper attenuated vaccine strains F and H

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Construction and biological activity of recombinant Newcastle disease LaSota attenuated vaccine expressing canine distemper virus fusion protein (F) or canine distemper virus hemagglutinin protein (H)

[0039] 1 Materials and methods

[0040] 1.1 Materials

[0041] 1.1.1 Cells, viruses, recombinant plasmids, chicken embryos and experimental animals: HEp-2 cells (ATCC CCL-23) and BHK-21 cells (ATCC CCL-10), the medium is DMEM containing 10% fetal bovine serum; Vero -E6 cells were preserved by our laboratory; recombinant poxvirus vTF7-3 (ATCC, VR-2153) stably expressing T7 polymerase was purchased from ATCC; NDV infectious clone plasmid pBRN-FL-PmeI and auxiliary nucleoprotein (pBS-NP) , phosphoprotein (pBS-P) and large polymerase protein (pBS-L) recombinant plasmids were constructed and preserved by our laboratory [6，7] , they can also be constructed according to the specification example 1 of ZL200510097997.8; recombinant Newcastle disease attenuated vaccine...

Embodiment 2

[0067] Example 2 Pathogenicity test of recombinant virus, immune protection test to mice and immune test to dogs

[0068] 1. Materials and methods

[0069] 1.1 Pathogenicity test of recombinant virus

[0070] According to O.I.E standard method [9] The pathogenicity indexes of rLa-CDVR-F and rLa-CDVR-HF F3 generations of recombinant viruses were measured, such as average chick embryo lethal time (MDT), intracerebral lethality index (ICPI) and intravenous pathogenicity index (IVPI).

[0071] 1.2 Immunization test of recombinant virus on mice

[0072] Divide 4-week-old female BALB / c mice into four groups, 15 in each group, recombinant strain rLa-CDVR-F, recombinant strain rLa-CDVR-H, rLa-CDVR-F / H mixed virus solution (1:1 ) by about 10 8.5 EID 50 / 100μL dose (half the egg infection dose (EID) 50 )) Intranasal drip and intramuscular injection were used to immunize each rat, 130 μL each. The fourth group was the normal control group, and the parental strain rLaSota was immuni...

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Abstract

The invention relates to a recombination newcastle disease LaSota attenuated vaccine for expressing canine distemper virus fusion protein (F) or canine distemper virus hemagglutinin protein (H). Particularly, the recombination newcastle disease LaSota attenuated vaccine is rLa-CDVR-F or rLa-CDVR-H. The invention further discloses a method for preparing the recombination newcastle disease LaSota attenuated vaccine and application of the recombination newcastle disease LaSota attenuated vaccine in preparation of vaccines/kits.

Description

technical field [0001] The present invention relates to the field of recombinant virus vaccines, more specifically, the present invention relates to a recombinant Newcastle disease LaSota attenuated vaccine expressing canine distemper virus fusion protein (F) or canine distemper virus hemagglutinin protein (H or HA), more specifically Specifically, the recombinant Newcastle disease LaSota attenuated vaccine is rLa-CDVR-F or rLa-CDVR-H. The invention also discloses a method for preparing the recombinant Newcastle disease LaSota attenuated vaccine and the application of the recombinant Newcastle disease LaSota attenuated vaccine in preparing vaccines / kits. Background technique [0002] Canine distemper (CD) is an acute, highly contagious infectious disease caused by canine distemper virus (CDV) infection. The disease can infect not only canids, but also mustelidae, procyonidae, cats and other animals. The incidence rate is high, almost 100%, and the clinical symptoms are dive...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/01A61K39/295A61K39/17A61K39/175A61P31/14C12N15/85C12R1/93
Inventor 步志高陈化兰葛金英夏咸柱王喜军高玉伟
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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