Duck hemorrhagic ovaritis virus strain, inactivated vaccine and preparation method thereof

A technology for hemorrhagic oophoritis and inactivated vaccines is applied in the field of virus strains and inactivated vaccines, duck hemorrhagic oophoritis virus strains, inactivated vaccines and their preparation fields, which can solve the problem that the vaccine evaluation model has not been established and restricts vaccines. development and other issues

Active Publication Date: 2013-04-17
INST OF ZOOTCHNICS & VERTERINARY SCI BEIJING ACAD OF AGRI & FORESTRY SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, due to the lack of better vaccine strains, large-scale and high-titer laboratory hosts for cult

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] a. Vaccination: the duck hemorrhagic oophoritis virus through the allantoic cavity according to 10 3 ELD 50 Inoculate 10-day-old sensitive duck embryos at a dose of 1 piece, and incubate at 37°C;

[0039] b. Harvesting: Harvest duck embryo bodies, chorioallantoic membranes, and allantoic fluid that died 48 to 120 hours after inoculation and had obvious disease scars, crushed at 20,000 r / min for 2 minutes, repeatedly frozen and thawed twice, and stored at -20°C. It is duck embryo tissue fluid; according to the method for determining the median lethal dose and the half infection dose specified in appendix 7 of "The Veterinary Pharmacopoeia of the People's Republic of China" (version three in 2010), the virus content assay is carried out, and the proliferating virus titer is 10 7.1 ELD 50 / 0.1ml;

[0040] c. Inactivation: Add formaldehyde solution with a mass concentration of 37% to the duck embryo tissue fluid to make the final content 0.2%, shake well and inactivate a...

Embodiment 2

[0049] a. inoculation: the duck hemorrhagic oophoritis virus cell adaptation virus is inoculated in the monolayer of duck embryo fibroblasts in a ratio of 0.1%;

[0050] b. Harvest: Harvest the cells when the cytopathic effect (CPE) reaches 80% to 90% 72 to 96 hours after inoculation, and put the sterile beaker containing the harvested cell pellet in an ice bath at 0°C with a cell lyser at 85 Under the condition of % power, lyse for 30s / time, lyse for 3 times, and freeze at -20°C to be the virus cell lysate; according to the median lethal dose, The virus content determination is carried out by the determination method of the half infection amount, and the proliferating virus titer is 10 6.5 TCID 50 / 0.1ml;

[0051] c. Inactivation: Add formaldehyde solution with a mass concentration of 37% to the virus cell lysate to make the final content reach 0.1%, shake well and inactivate at 4°C for 7 days;

[0052] d emulsification:

[0053] Preparation of aqueous phase: Centrifuge t...

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Abstract

The invention discloses a duck hemorrhagic ovaritis virus strain, an inactivated vaccine and a preparation method thereof. The duck hemorrhagic ovaritis virus is named duck hemorrhagic ovaritis flavivirus HB strain, and the collection number of the duck hemorrhagic ovaritis flavivirus HB strain is CCTCC V201122. The virus is ssRNA virus with cyst membrane; virions are basically spherical, and the diameter is 40 to 60 nanometers; the virus is sensitive to ether and chloroform; most detergents can quickly inactivate the virus; the virus can also be inactivated when fumigated in 0.1-percent formalin for 6 hours at 37 DEG C; and the duck hemorrhagic ovaritis virus cannot agglutinate red blood cells of chickens, ducks and turkeys, but can agglutinate 1-percent red blood cells of doves. The inactivated vaccine is prepared by inoculating the duck hemorrhagic ovaritis virus, harvesting, inactivating, concentrating and emulsifying. The vaccine has the advantages of high safety, quick immune response, long immune duration and the like.

Description

technical field [0001] The invention relates to a virus strain and an inactivated vaccine, in particular to a duck hemorrhagic oophoritis virus strain, an inactivated vaccine and a preparation method thereof, belonging to the technical field of bioengineering. Background technique [0002] Duck hemorrhagic oophoritis (Duck Hemorrhagic, DH) is a newly popular, acute and highly contagious infectious disease of ducks. Its pathogen is Duck Hemorrhagic Ovaritis Virus (DHOV). Belonging to members of the Flaviviridae, it infects ducks of different ages, causing duck ovarian bleeding, decreased egg production, atrophy of ovaries, testes, oviducts and vas deferens, and increased mortality of commercial meat ducks. [0003] Since the disease occurred in Guangdong, Guangxi, Fujian, Jiangxi, Zhejiang, Anhui, Jiangsu, Shandong, Hebei and Beijing in June 2010, duck hemorrhagic oophoritis has become a major epidemic that recently endangered the duck industry in China. one. The virus main...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12R1/93
Inventor 刘月焕杨保收韩春华何平有林健郁宏伟徐倩倩潘洁梁武
Owner INST OF ZOOTCHNICS & VERTERINARY SCI BEIJING ACAD OF AGRI & FORESTRY SCI
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