Inactivated vaccine for preventing and controlling new chicken reovirus and preparation method thereof

A technology of reovirus and inactivated vaccines, which is applied in the field of inactivated vaccines and preparations for the prevention and treatment of new chicken reoviruses, can solve the problems of ineffective antiviral and antibacterial treatment methods, and can not be prevented and controlled, so as to prevent and control infection and outbreak, simplify the production process, and realize the effect of large-scale production

Active Publication Date: 2018-08-10
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The study found that the outbreak of the disease was caused by a new type of chicken reovirus infection. At present, there is no vaccine to prevent it. The exist

Method used

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  • Inactivated vaccine for preventing and controlling new chicken reovirus and preparation method thereof
  • Inactivated vaccine for preventing and controlling new chicken reovirus and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0036] In the preparation method of the above-mentioned inactivated vaccine, the LMH cell line is used to culture the virus. The cell line has a very good susceptibility to the novel chicken reovirus of the present invention. It can be continuously passaged, and can also be cultured in suspension with good growth. Conducive to virus proliferation and vaccine production.

[0037] Adjuvants refer to substances that can effectively promote, enhance or prolong the body's specific immune response to antigens after being added to vaccine preparations. The choice of adjuvant is the basis of the vaccine emulsification process. A stable vaccine emulsion can effectively improve the ability of animals to resist diseases. Therefore, choosing a suitable adjuvant and determining the best ratio of water to oil are of great significance to the stability of the vaccine.

Embodiment 1

[0040] Example 1: Discovery and identification of new chicken reovirus

[0041] 1. Virus isolation:

[0042] (1) For arthritic chicks that were clinically identified as positive by PCR reaction, the swollen tendon tissue was collected by necropsy and placed in a 15mL centrifuge tube, and 5 times the volume of serum-free DMEM medium was added, homogenized and repeatedly frozen and thawed. 3 times, after each thawing, shake on the shaker for 1-2 minutes, and then perform the next freeze-thaw; after freezing and thawing, centrifuge the 15mL centrifuge tube containing the sample in the centrifuge at 4000rpm for 5min; take the supernatant, 0.22μm micropores After the filter membrane is filtered, spare;

[0043] (2) According to the proliferation characteristics of reo, chicken liver cancer cells (LMH) were selected for virus isolation. According to the conventional cell culture method, wait for 25cm 2 When the cells in the cell flask are covered with a monolayer, aspirate the culture me...

Embodiment 2

[0059] Example 2: Preparation of inactivated vaccine

[0060] (1) The propagation and harvest of viruses:

[0061] The isolated and identified new chicken reovirus is inoculated into a well-growing LMH cell line, and the virus is proliferated in large quantities to obtain sufficient virus liquid. The obtained sufficient cell virus liquid was frozen at -20°C, and the cell virus liquid was collected after two freeze-thaw cycles.

[0062] (2) Purification of virus:

[0063] Use established PCR, RT-PCR and other detection methods to detect whether the obtained virus liquid contains other common viruses. Test items include avian influenza virus (AIV), Newcastle disease virus (NDV), fowlpox virus (APV), Mycoplasma synovium (MS), infectious bronchitis virus (IBV) and infectious bursal disease virus (IBDV) Etc.; Centrifuge the harvested virus solution at 5000r / min for 5min to remove cell debris and other impurities, then use ultracentrifugation at 60000r / min for 2h, and resuspend with DMEM ...

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Abstract

The invention discloses an inactivated vaccine for preventing and controlling a new chicken reovirus and a preparation method thereof. The inactivated vaccine contains inactivated chicken reovirus strain virus liquor with the effective dose on prevention or treatment. The preservation number of a chicken reovirus strain is CCTCC NO:V201817. The prepared inactivated vaccine for preventing and controlling the new chicken reovirus is good in safety, no any local or whole-body adverse reactions caused by the vaccine are generated, and detection of all indexes is stable and effective. After broilers are immunized by the inactivated vaccine, the broilers can obtain the high antibody level, the duration is long, infection and outburst of the new chicken reovirus can be pertinently prevented and controlled, and effective immunoprotection is provided for chicken flocks. Meanwhile, the production process of the vaccine is simplified, and large-scale production can be achieved.

Description

Technical field [0001] The invention relates to the technical field of veterinary biological products, in particular to an inactivated vaccine for preventing and treating new chicken reovirus and a preparation method thereof. Background technique [0002] Avian orthoreovirus (Avian orthoreovirus, ARV) belongs to the Reoviridae family and the genus Orthoreovirus. It is the pathogen that causes viral arthritis in chickens. It mainly infects broiler chickens, but also chickens with both meat and eggs. ARV mainly affects chicken's tarsal joints, toe joints, tibial and metatarsal joints and their tendons, causing joint synovitis or tenosynovitis. Diseased chickens usually show symptoms of limp, squatting, unwillingness to walk, loss of appetite, or even abolition, resulting in low feed conversion rate, increasing number of dead chickens, reducing production efficiency, and causing serious economic losses to the poultry industry. Since the disease was first reported in my country in t...

Claims

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Application Information

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IPC IPC(8): A61K39/15A61P31/14C12N7/00C12R1/93
CPCA61K39/12A61K2039/5252A61K2039/552A61K2039/575A61P31/14C12N7/00C12N2720/12221C12N2720/12234
Inventor 刁有祥唐熠姜晓宁
Owner SHANDONG AGRICULTURAL UNIVERSITY
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