H9 subtype avian influenza virus strain

An avian influenza virus and virus technology, applied in the direction of antiviral agents, viruses/phages, biochemical equipment and methods, etc., can solve problems such as poor immune effect of vaccines, improve uniformity, improve antibody levels, and improve safety. Effect

Active Publication Date: 2015-09-30
YEBIO BIOENG OF QINGDAO
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

[0004] The purpose of the present invention is to provide a kind of H9 subtype avian influenza virus strain; And use this virus strain to prepar...

Method used

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  • H9 subtype avian influenza virus strain
  • H9 subtype avian influenza virus strain

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Embodiment 1

[0013] Embodiment 1, the screening of antigenic strain (H9 subtype avian influenza virus QDY strain) for making seedlings

[0014] In 2013, from the infected chickens that had been vaccinated with the H9 subtype avian influenza virus vaccine in Chengyang, Shandong, aseptically collected heart, spleen and other organs, larynx, and cloacal swabs of the affected chickens, and put them in a sterile plate under sterile conditions. After crushing, add physiological saline containing 4000 units / ml double antibody at a ratio of 1:5, and freeze and thaw repeatedly at -20°C for 3 times. Put the cotton swab directly into the physiological saline containing 10,000 units of double antibody, act for 4 hours at 2-8°C, centrifuge the viscera and cotton swab at 3500 rpm for 10 minutes, take the supernatant, mix and inoculate 10-day-old SPF chicken embryo 5 Pieces, 0.2ml / piece. Chicken embryos were incubated at 37°C, inspected once a day in the morning and evening, and the dead embryos were ta...

Embodiment 2

[0018] Embodiment 2: Preparation of antigen for seedling production

[0019] 1. Preparation of virus liquid for seedling production Inoculate 10-11-day-old SPF chicken embryos with virus seeds (QDY strain) for production through the allantoic cavity, 0.2ml per embryo, incubate at 36°C-37°C, and irradiate the embryos twice a day, Take the chicken embryos that died after 24 hours, harvest them at 96 hours regardless of whether they are dead, cool them at 4-8°C for 12-24 hours, harvest the chicken embryo liquid, mix them in a sterile container, and store them at 2-8°C.

[0020] 2. Preparation of chicken α-interferon protein for seedling production Fermentation tank aeration culture, prepare medium according to 70% of the volume of the fermentation tank, adjust the pH value to 7.0-7.2 with 1mol / L NaOH, add defoaming according to 0.02% of the total volume of the medium agent. After sterilization, inoculate the strains for production with 1% to 2% inoculation amount, add 10% kanamy...

Embodiment 3

[0028] One, the preparation of inactivated vaccine: the semi-finished product antigen after passing the inspection is carried out vaccine preparation (in the following preparation, each liquid component is calculated by volume ratio).

[0029] (1) Oil phase preparation Take 95 parts of veterinary white oil and 1 part of aluminum stearate, put them in the oil phase preparation tank and heat to 80°C, then add 805 parts of Siben-805, and when the temperature reaches 115°C, keep 30min, cool down and set aside.

[0030] (2) Preparation of aqueous phase Dilute chicken interferon-alpha protein with normal saline to 2×10 4.0 ~2×10 6.0 Unit / 0.1ml; Dilute the inactivated H9 subtype avian influenza virus solution with normal saline to not less than 2×10 6.0 EID 50 / 0.1ml. Take chicken α-interferon protein liquid and inactivated H9 subtype avian influenza virus liquid respectively, and mix them to prepare antigenic liquid for vaccine preparation. Take 5 parts of sterilized Tween-80 a...

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Abstract

The invention aims at providing an H9 subtype avian influenza virus strain. Vaccine is prepared by the virus strain to solve the problem that existing H9 subtype avian influenza virus vaccine is poor in immune effect caused by a novel virus. The H9 subtype avian influenza virus QDY strain (Avian Influenza Virus) is preserved in the typical culture preservation center in China in Wuhan University on April 29, 2015, and the preservation number is CCTCC NO: V201517. After the H9 subtype avian influenza virus (QDY strain) is inoculated to a chick embryo, a virus solution is collected, after ultrafiltration concentration and formaldehyde solution inactivation are performed, oil adjuvant is added, and then mixing and emulsifying are performed to prepare the vaccine. According to the prepared vaccine, the antibody level after immunization can be improved, the antibody uniformity after immunization can be improved, and the immune effect of the vaccine can be guaranteed; the vaccine has the advantages of being efficient and good in safety.

Description

technical field [0001] The invention belongs to the technical field of biological veterinary medicine, and in particular relates to an H9 subtype avian influenza virus strain. Background technique [0002] H9 subtype avian influenza is a low-pathogenic disease, but mixed infection with other pathogens, especially Escherichia coli, can increase its pathogenicity, cause a serious decline in egg production and a significant increase in mortality, and bring serious harm to the poultry industry. The existing avian influenza (subtype H9) vaccine has been widely used clinically, but there are problems of low antibody level and antibody uniformity, which affect the prevention and control effect of the vaccine. [0003] The occurrence of low pathogenic H9 subtype avian influenza is potential, but it can also bring disaster to the poultry industry, and the threat to humans is equally serious. The epidemic characteristics of low pathogenic avian influenza virus are extremely wide dist...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/145A61P31/16C12R1/93
Inventor 刘新文杜元钊宫晓李陆梅程增青邹敏王龙
Owner YEBIO BIOENG OF QINGDAO
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