Chicken Newcastle Disease Virus Attenuated Strain, Inactivated Vaccine and Its Application

A technology for chicken Newcastle disease virus and chicken Newcastle disease, which is applied to attenuated strains of chicken Newcastle disease virus, inactivated vaccines and their application fields, and can solve the problems of virus hemagglutination spectrum, differences in virus thermal stability and the like

Inactive Publication Date: 2016-04-06
NORTHEAST FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

NDV has only one serotype, but the pathogenicity, biological characteristics and physicochemical characteristics of different strains are not the same. difference

Method used

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  • Chicken Newcastle Disease Virus Attenuated Strain, Inactivated Vaccine and Its Application
  • Chicken Newcastle Disease Virus Attenuated Strain, Inactivated Vaccine and Its Application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Isolation, cultivation and identification of chicken Newcastle disease virus attenuated strain (DL11 strain) of embodiment 1

[0045]1. Collection of disease materials

[0046] Chicken disease materials come from a farm in the suburbs of Daqing City, Heilongjiang Province. The livers of dying chickens are collected. After grinding, they are treated with 1,500 units of penicillin and streptomycin solution at 4°C for 12 hours, centrifuged at 2,000 r / min, and the supernatant is taken to inoculate chicken embryos and collected for 48 hours. 96h dead chicken embryo allantoic fluid.

[0047] 2. Hemagglutination (HA) and hemagglutination inhibition (HI) tests of virus isolates

[0048] Hemagglutination test was carried out on the harvested chicken embryo fluid, and the hemagglutination value of HA was 8log 2 .

[0049] HI test was carried out with avian influenza H5, H7 and H9 subtype standard positive sera, ND and EDS standard positive sera and chicken embryo allantoic flu...

Embodiment 2

[0073] The preparation of embodiment 2 chicken Newcastle disease virus DL11 strain seed virus

[0074] 1. Establishment of basic seed batches

[0075] Use the screened 20th generation seed virus (CGMCC NO.7958) to inoculate the CEF grown into a monolayer, and after adsorption at 37°C for 1 hour, add DMEM containing 2% calf serum, and cool to 37°C, 5% CO 2 Environmental culture, after 80% of the cells have lesions, freeze and thaw three times at -20°C, harvest the cytotoxicity, and harvest after two freeze-thaws. Continuous passage does not exceed 3 generations.

[0076] 2. Production seed batch establishment

[0077] Take the basic seeds, dilute them 10,000 times with sterilized normal saline, insert 1% of the virus culture solution into the monolayer cell culture, absorb at 37°C for 1 hour, add DMEM containing 2% calf serum, to 37°C, 5% CO 2 Environmental culture, after 80% of the cells have lesions, freeze and thaw 3 times at -20°C, and harvest the toxic cell culture med...

Embodiment 3

[0078] The preparation of embodiment 3 chicken Newcastle disease inactivated vaccines

[0079] In order to verify the characteristics of cytotoxicity, 10 batches of inactivated vaccines were prepared with the same method using the screened 20th-generation seed virus (CGMCC NO.7958), and the relevant technical indicators were determined, as described below.

[0080] 1. Preparation of virus liquid

[0081] 1.1 Preparation of cells

[0082] Take SPF chicken embryos aged 9-11 days, remove the head, limbs and viscera, and cut them into 1mm 3 Tissue blocks left and right. Wash 3 times with PBS, digest with 0.25% trypsin for 5-15min, discard the trypsin, add DMEM containing 10% calf serum, blow the cells evenly with a pipette, put them into a culture bottle, and culture them at 37°C until Cells were used after growing into a monolayer.

[0083] 1.2 Inoculation and harvest

[0084] Wash the monolayered CEF with PBS for 3 times, inoculate the 20th generation seed virus (CGMCC NO.7...

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Abstract

The invention discloses an attenuated strain and an inactivated vaccine of the Newcastle disease virus and application thereof, belonging to the field of separation and application of a Newcastle disease virus strain. The Newcastle disease virus attenuated strain (DL11 strain) is separated from chicken disease materials and has a microbe preservation number of CGMCC No. 7958. According to results of hemagglutination and hemagglutination inhibition tests, the separated virus strain is the Newcastle disease virus strain; and intravenous pathogenicity index determination tests prove that the virus strain is the Newcastle disease virus attenuated strain. The erythrocyte agglutination value and virus content in a semi-finished product of the separated Newcastle disease virus attenuated strain are both higher than standards prescribed in Veterinary Biological Product Regulations; and the separated Newcastle disease virus attenuated strain has low preparation cost and meets requirements for mass production. The inactivated vaccine prepared from the attenuated strain has high immune efficacy, can resist attack by a virulent strain and is applicable as a vaccine strain for prevention or diagnosis of the Newcastle disease virus.

Description

technical field [0001] The present invention relates to an attenuated strain of virus, in particular to an attenuated strain of Newcastle disease virus (Newcastle disease virus), the invention also relates to an inactivated vaccine prepared by the attenuated strain of Newcastle disease virus and its use in the preparation of drugs or reagents for preventing, treating or diagnosing Newcastle disease The invention belongs to the field of separation and application of chicken Newcastle disease virus attenuated strain. Background technique [0002] Newcastle disease (Newcastle disease, ND) is an acute, highly contagious disease of poultry, and its pathogen is Newcastle disease virus (Newcastled disease virus, NDV). ND poses a serious threat to the world's poultry industry and causes huge losses. It is designated as a Class I severe infectious disease by the International Office of Epizootics, so Newcastle disease is the focus of poultry disease research. Although Newcastle dise...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N7/00A61K39/17A61P31/14C12R1/93
Inventor 邢明伟柴洪亮曾祥伟侯志军田丽红
Owner NORTHEAST FORESTRY UNIVERSITY
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