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Preparation method of duck reovirus inactivated vaccine

A technology of reovirus and inactivated vaccine, which is applied in the field of preparation of duck reovirus inactivated vaccine, can solve the problem of uncontrollable epidemic situation, etc. Effect

Active Publication Date: 2019-10-22
广东渔跃生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Then, although the multiple selection vaccines that have been released at present, the situation that the epidemic situation cannot be controlled still occurs. Therefore, it is still a difficult problem to be solved urgently to research and develop a duck reovirus inactivated vaccine with better suppression effect.

Method used

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  • Preparation method of duck reovirus inactivated vaccine
  • Preparation method of duck reovirus inactivated vaccine
  • Preparation method of duck reovirus inactivated vaccine

Examples

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Effect test

Embodiment 1

[0036] Embodiment 1, a kind of preparation method of duck reovirus inactivated vaccine

[0037] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and use DMEM / F12 culture medium containing 5% newborn bovine serum and 1000 units / mL penicillin-streptomycin double antibody at 37°C, 5% CO 2 Cultured under the conditions of 2 days, then washed the cells twice with serum-free DMEM / F12 culture medium to obtain cell carriers; the LMH cells were cultured in spinner bottles or cultured in microcarrier reactors, and the amount of microcarriers used was 4 g / Lift;

[0038] Inoculation of S2 virus: inoculate the duck reovirus liquid into the cell carrier obtained in step S1 according to the final volume of 1:1000, place it at 37°C for 30 minutes, absorb the virus liquid, then add medium A, and incubate at 37°C , 5%CO 2 48h under the condition of culturing, 80% cytopathy occurs, and diseased cells are obtained;

[0039] Described culture medium A comprises DMEM / ...

Embodiment 2

[0047] Embodiment 2, a kind of preparation method of duck reovirus inactivated vaccine

[0048] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and use DMEM / F12 culture medium containing 8% newborn bovine serum and 1500 units / mL penicillin-streptomycin double antibody at 37°C, 5% CO 2 Cultivate under the condition of 2 days, then wash cells 3 times with serum-free DMEM / F12 culture medium, obtain cell carrier; The described LMH cell is placed in the culture of microcarrier reactor, and microcarrier usage amount is 6 grams / liter;

[0049] Inoculation of S2 virus: Inoculate the duck reovirus liquid into the cell carrier obtained in step S1 according to the final volume of 1:2000, place it at 37°C for 40 minutes, then absorb the virus liquid, then add medium A, and incubate at 37°C , 5%CO 2 Under the conditions of 56 hours, 80% of the cells were damaged, and the diseased cells were obtained;

[0050] Described culture medium A comprises DMEM / F12 basal...

Embodiment 3

[0058] Embodiment 3, a kind of preparation method of duck reovirus inactivated vaccine

[0059] Preparation of S1 cell carrier: Digest and disperse LMH cells with trypsin, and use DMEM / F12 culture medium containing 10% newborn bovine serum and 2000 units / mL penicillin-streptomycin double antibody at 37°C, 5% CO 2 Cultivate under the condition of 3 days, then wash cells 3 times with serum-free DMEM / F12 culture medium, obtain cell carrier; Described LMH cell is placed in the culture of microcarrier reactor, and microcarrier usage amount is 8 grams / liter;

[0060] Inoculation of S2 virus: inoculate the duck reovirus liquid into the cell carrier obtained in step S1 according to the final volume of 1:4000, place it at 37°C for 60 minutes, then absorb the virus liquid, then add medium A, and incubate at 37°C , 5%CO 2 72h under the condition of culture, 80% cytopathy occurs, and diseased cells are obtained;

[0061] Described culture medium A comprises DMEM / F12 basal medium, also c...

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Abstract

The invention belongs to the technical field of veterinary biological products, and particularly relates to a preparation method of a duck reovirus inactivated vaccine. In the invention, an LMH passage cell line is used as a carrier cell for virus proliferation; a culture medium A consisting of lysine, recombinant human insulin, human serum albumin, protamine, biotin, polysaccharide sulfate and agrowth factor is used for culture; and the virus fluid is collected and inactivated to prepare a vaccine. The LMH passage cell line is used as a carrier cell for virus proliferation without the need of adding pancreatin, and the LMH cells have a clear background, have no exogenous pathogens and are easy for proliferation, which can effectively simplify the process and reduce costs. Meanwhile, theduck reovirus inactivated vaccine prepared by the invention has high virus content, good stability and high safety, and is an ideal duck reovirus inactivated vaccine.

Description

technical field [0001] The invention belongs to the technical field of veterinary biological products, and in particular relates to a preparation method of duck reovirus inactivated vaccine. Background technique [0002] Duck reovirus is a new disease that has emerged in my country in recent years. It is characterized by irregular liver necrosis, hemorrhagic spots, cardiac muscle and supracavitary bursa hemorrhage. The disease is caused by a new RNA virus, and its pathogen is Reoviridae Orthoreovirus is a new type of duck reovirus. The morbidity and mortality vary greatly, but the younger the sick duck, the higher the morbidity and mortality. According to reports by Cheng Anchun et al., infected ducks aged 3 to 50 days, including Peking Duck, Cherry Valley Duck, Tianfu Meat Duck, Sichuan Shelduck, etc.; the incubation period is 4 to 6 days; , with the development of the disease, lying on the ground, drinking water, diarrhea, dyspnea, eyelid congestion, bleeding and severe s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/15A61P31/14C12N5/09C12N7/00C12N7/02
CPCA61K39/12A61P31/14C12N5/0693C12N7/00C12N2720/12234A61K2039/5252C12N2531/00C12N2720/12251Y02A50/30
Inventor 张毓金严悌昆谢秉超黄淑芬张桂平
Owner 广东渔跃生物技术有限公司
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