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Goose paramyxovirus chicken embryo fibroblast cell adapted strain and application thereof

A goose paramyxovirus and fibroblast technology, applied in antiviral agents, viruses/bacteriophages, medical preparations containing active ingredients, etc., can solve the problems of poor product quality between batches and high cost, and achieve stable product quality, Good immunogenicity and high antibody titer

Active Publication Date: 2015-04-08
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

At present, most of the goose paramyxovirus vaccines on the market are tissue seedlings produced from chicken embryos. Compared with cell seedlings, they have disadvantages such as high cost and large batch-to-batch variation in product quality.

Method used

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  • Goose paramyxovirus chicken embryo fibroblast cell adapted strain and application thereof
  • Goose paramyxovirus chicken embryo fibroblast cell adapted strain and application thereof
  • Goose paramyxovirus chicken embryo fibroblast cell adapted strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Isolation and Identification of Goose Paramyxovirus

[0030] 1. Experimental method

[0031] 1.1 Isolation of virus

[0032] The disease material came from a farm in the suburbs of Daqing City, Heilongjiang Province. The liver, spleen and other disease materials of the dead goose were aseptically collected, mixed, chopped and ground, and sterilized PBS with a double antibody content of 2000IU / mL was added at a ratio of 1:3 (v / v) , placed in a refrigerator at 4°C for 4 hours, and centrifuged at 4000r / min for 5 minutes; take the supernatant and inoculate five 10-day-old SPF chicken embryos through the allantoic cavity, 0.2mL / piece, and incubate at 37°C for 120 hours; discard the dead chicken embryos within 24 hours, After that, the eggs were illuminated twice a day, and the dead chicken embryos were stored in a refrigerator at 4°C for 4-24 hours; the allantoic fluid and amniotic fluid of the dead chicken embryos were collected from 48h to 120h, and used as the ...

Embodiment 2

[0079] Embodiment 2 animal immune protection test

[0080] 1. Experimental method

[0081] 1.1 Preparation of oil emulsion inactivated vaccine

[0082] 1.1.1 Preparation of virus solution

[0083] The goose paramyxovirus DQ01 strain, DQ02 strain, DQ03 strain, DQ04 strain and DQ05 strain isolated in Example 1 were cultured by chicken embryo fibroblast spinner bottles respectively, and the cell culture virus liquid was collected to measure HA and TCID 50 In addition, the original strain that embodiment 1 isolates is inoculated into SPF chicken embryo (collects allantoic fluid), measures its HA and EID 50 .

[0084] 1.1.2 Inactivation and inactivation test of virus liquid

[0085] 1.1.2.1 Inactivation of virus fluid

[0086] Add the tested virus liquid to a final concentration of 0.1% formaldehyde solution for inactivation, and inactivate at 37°C for 24 hours.

[0087] 1.1.2.2 Inactivation test

[0088] (1) Inactivation test of cell virus liquid

[0089] Take the inactiva...

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Abstract

The invention discloses a goose paramyxovirus chicken embryo fibroblast cell adapted strain and application thereof and belongs to the field of separation and application of goose paramyxovirus. According to the invention, a goose paramyxovirus chicken embryo fibroblast cell adapted strain DQ03 with the microbial preservation number of CGMCC No.9901 is separated firstly. The invention also discloses a method for preparing a goose paramyxovirus preventing inactivated vaccine. The method for preparing the goose paramyxovirus preventing inactivated vaccine comprises the following steps: (1) cultivating the goose paramyxovirus chicken embryo fibroblast cell adapted strain DQ03, and collecting virus bulk; (2) inactivating the virus bulk; and (3) centrifuging, taking virus supernate, mixing with adjuvant, and emulsifying, thus obtaining the goose paramyxovirus preventing inactivated vaccine. The separated goose paramyxovirus chicken embryo fibroblast cell adapted strain DQ03 is highly adapted to chicken ambryo fibroblast cells and is applicable to cell culture; the immunogenicity is good, and the goose paramyxovirus chicken embryo fibroblast cell adapted strain DQ03 has the advantages of high titer of a produced antibody, low production cost and the like compared with a vaccine prepared from embryo virus.

Description

technical field [0001] The present invention relates to goose paramyxovirus, in particular to an isolated goose paramyxovirus chicken embryo fibroblast-adapted strain, and the present invention also relates to the preparation of the isolated goose paramyxovirus chicken embryo fibroblast-adapted strain The invention relates to a vaccine for myxovirus disease, and belongs to the field of separation and application of goose paramyxovirus. Background technique [0002] Goose paramyxovirus (GPMV) is a member of the Paramyxoviridae family, Paramyxoviridae subfamily, and the genus Mumps virus. Goose paramyxovirus disease is a severe infectious disease with high morbidity and mortality mainly characterized by digestive tract lesions. It can occur all year round, with an incidence rate of 40%-100%, an average of about 60%, and a mortality rate of 40%. It is 30%-100%, and the average is about 40%. Geese of different ages are susceptible to infection, but the younger the age, the hig...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K39/155A61P31/14C12R1/93
Inventor 孙德君丁国杰杨万秋梁宛楠李来旭宋扬藏玉婷刘鑫莹张智明李鑫窦海艳
Owner 哈药集团生物疫苗有限公司
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