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Duck hemorrhagic oophoritis inactivated vaccine production method by using cell line and product thereof

A technology for hemorrhagic oophoritis and inactivated vaccines, which is applied in the field of vaccine preparation, can solve problems such as the establishment of vaccine evaluation models, the failure to find vaccine strains, and restrictions on vaccine development, and achieve good economic benefits and application prospects. Pollution probability is small, the production process is simple and stable

Active Publication Date: 2013-07-17
RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The effective measure recommended by experts to prevent and treat the disease is to inject vaccines. However, no good vaccine strains, large-scale and high-titer cell lines for culturing the virus have yet been found, and the vaccine evaluation model has not yet been established, which restricts the development of current vaccines.

Method used

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  • Duck hemorrhagic oophoritis inactivated vaccine production method by using cell line and product thereof
  • Duck hemorrhagic oophoritis inactivated vaccine production method by using cell line and product thereof
  • Duck hemorrhagic oophoritis inactivated vaccine production method by using cell line and product thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1 Screening of virus-adapted cell lines

[0032] According to the characteristics of the virus, four cell lines, bovine embryonic kidney cell line (MDBK), hamster kidney cell line (BHK21), pig kidney cell line (PK15), and African green monkey kidney cell line (Vero), were selected for duck hemorrhagic oophoritis virus. Subculture of HB strain and selection of adaptive cell lines for duck hemorrhagic oophoritis virus.

[0033] All four cells were purchased from China Veterinary Drug Administration.

[0034] Duck hemorrhagic oophoritis virus HB strain, the preservation number is CCTCC V201122, was preserved in China Center for Type Culture Collection on July 1, 2011. The virus is an ssRNA virus with an envelope; the virus particles are roughly spherical, with a diameter of 40-60nm; the virus has strong resistance to the external environment, and it can be stored at 4°C for several weeks and stored at -20°C for several months , its infectivity is not affected;...

Embodiment 2

[0042] Example 2 Breeding of Duck Hemorrhagic Ovarian Virus Strain and Establishment of Virus Seed Batches

[0043] 1 Test method

[0044] 1.1 Breeding of strains

[0045]1.1.1 Virus purification: The duck embryo-adapted strain of duck hemorrhagic oophoritis virus HB strain was successively passaged on hamster kidney cell line (BHK21) and African green monkey kidney cell line (Vero) respectively for 3 times, and the cells were taken to subculture the adapted strain It was purified and cultured using 0.1% neutral red plaque purification technology, continuously purified three times, and the potency was determined;

[0046] 1.1.2 Virus inactivation: Take the cytovenom of the purified virus strains, freeze and thaw twice, add 0.1% formaldehyde of the total amount of cytovenom (V / V) to inactivate, and inactivate by shaking at 37°C for 24 hours;

[0047] 1.1.3 Washing and concentration of venom: The inactivated cell venom is concentrated by ultrafiltration through the Millipo...

Embodiment 3

[0061] Example 3 Preparation of Inactivated Vaccine of Duck Hemorrhagic Ovariitis Using Cell Line

[0062] 1 Test material

[0063] Cell line: hamster kidney cells (BHK21);

[0064] Seed virus: duck hemorrhagic oophoritis virus B propagated in embodiment 2 13 batch;

[0065] Cell growth solution: the composition is 90% DMEM solution by volume, 10% newborn bovine serum, and the pH value is 7.2;

[0066] Cell maintenance solution: the composition is 97% by volume DMEM solution, 3% newborn bovine serum, and the pH value is 7.3.

[0067] 2 Test method

[0068] 2.1 Subculture of cells

[0069] Hamster kidney cells (BHK21) were digested with EDTA-trypsin cell dispersion solution, added cell growth medium, continued to culture at 37°C, and waited for the cells to grow into a good monolayer before use;

[0070] 2.2 Virus inoculation and harvest

[0071] Take the BHK21 cell culture flask that has formed a good monolayer, discard the cell growth medium, and inoculate duck hemor...

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Abstract

A duck hemorrhagic oophoritis inactivated vaccine production method by using a cell line and a product thereof belong to the field of biotechnology. According to the method provided by the invention, the cell line is selected from hamster kidney cell line or African green monkey kidney cell line, and the preparation of the vaccine is accomplished by the following steps of: (1) breeding strain, (2) establishing virus seed group, (3) preparing cell venom, (4) inactivating virus, (5) washing and concentrating cell venom, (6) emulsifying and the like. The method provided by the invention has characteristics of simple and stable production process, easy operation and high security. The prepared duck hemorrhagic oophoritis inactivated vaccine has advantages of high quality, high yield, low cost, small inter-batch difference, good safety and high immune efficacy.

Description

technical field [0001] The invention relates to a preparation method of a vaccine and its product, more specifically to a method for producing an inactivated duck hemorrhagic oophoritis vaccine with a cell line and its product, belonging to the field of biotechnology. Background technique [0002] Duck Hemorrhagic Ovaritis (DHO) is a newly popular, acute and highly contagious infectious disease of ducks. Its pathogen is Duck Hemorrhagic Ovaritis Virus (DHOV). A member of the Flaviviridae family. The virus infected ducks of different ages, and the laying ducks mainly showed a sharp drop in feed intake, a sharp drop in egg production rate, and some ducks were paralyzed. Anatomical lesions were mainly manifested as ovarian hemorrhage, follicle deformation and degeneration, atrophy of ovary, yolk, testis, fallopian tube, and vas deferens, and miliary nodules on the surface of the liver. Commercial meat ducks show an increase in the dead rate. [0003] Recent scientific resear...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/12C12N7/08A61P31/14
Inventor 杨保收刘月焕何平有徐倩倩毛雅元韩春华林健刘涛
Owner RINGPU (BAODING) BIOLOGICAL PHARMACEUTICAL CO LTD
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