Type B acian metapneumovirus subunit vaccine

An avian metapneumovirus and subunit vaccine technology, which can be applied in vaccines, viruses, antiviral agents, etc., can solve the problem of high safety, and achieve the effects of good safety, improved antibody level, and improved uniformity

Active Publication Date: 2016-07-20
YEBIO BIOENG OF QINGDAO
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Type B acian metapneumovirus subunit vaccine
  • Type B acian metapneumovirus subunit vaccine
  • Type B acian metapneumovirus subunit vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Amplification and sequence analysis of F protein

[0022] In 2010, symptoms of swollen head syndrome appeared in several breeding chicken farms in Shandong Province, and the affected chickens had been injected with the existing avian pneumovirus vaccine before, and it was speculated that the infected virus had mutated; Screening for lung viruses. Finally, an avian pneumovirus SHS / A3 was screened out.

[0023] In order to verify the antigenicity of the screened virus, virus strains from 5 different sources, including the screened strain SHS / A3, were used as antigens to prepare vaccines. After immunizing SPF chickens, the SHS / A3 strain virus solution was used to challenge the virus. The results It shows that compared with other poultry pneumovirus vaccines; the vaccine prepared by itself has a better immune effect (p<0.05); therefore, it is determined that genetic variation has occurred.

[0024] 1. Amplification of Type B Avian Metapneumovirus SHS / A3 Strain ...

Embodiment 2

[0035] Embodiment 2: Recombinant expression of F protein

[0036] 1. The preparation method of recombinant type B avian metapneumovirus F protein

[0037] The method comprises the following steps: a. constructing an expression vector; b. constructing an expression strain; c. inducing, extracting and purifying the recombinant F protein.

[0038] a. Construct expression vector:

[0039] The positive clone plasmid pMD18-T-F and the expression vector pPICZα vector were double-digested with KpnI and NotI respectively, electrophoresed on a 1.2% agarose gel, and then recovered with a DNA gel recovery kit to obtain about 1.6kb and 3.3kb fragments, respectively. The pPICZα-F expression vector was constructed by directional ligation at 16°C ( Figure 4 It is the enzyme digestion identification diagram of the high-efficiency expression vector of the embodiment of the present invention); after sequencing and verifying that the sequence and reading frame are correct, the plasmid is linea...

Embodiment 3

[0044] Embodiment 3: the preparation of subunit vaccine

[0045] 1. Subunit vaccine preparation

[0046] 1. Preparation of bacterial solution for making seedlings Inoculate X33-F strains in YPD liquid medium containing bleomycin, and shake and culture at 30°C for 16-18 hours. Then streak and inoculate in YPD solid medium with bleomycin, select 2 to 3 typical colonies and mix them in a small amount of YPD liquid medium, place them in a shaker at 30°C for 18 hours, and quantitatively pack them. After inspection, it is used as a first-class seed. Take the first-grade seeds and inoculate them in BMGY liquid medium, culture them with shaking at 30°C for 16-18 hours, and store them at 2-8°C after microscopic examination

[0047] 2. Preparation of protein for seedling production. Add BMGY incomplete liquid medium according to 60% (V / V) of the volume of the fermenter, and add antifoaming agent according to 0.1% (V / V) of the medium, and pass through high-temperature steam for 30 minute...

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Abstract

The invention provides a type B acian metapneumovirus subunit vaccine.The type B acian metapneumovirus subunit vaccine is prepared from an antigen and a vaccine adjuvant, wherein inactivated type B acian metapneumovirus F protein serves as the antigen, and the amino acid sequence of the type B acian metapneumovirus F protein is SEQ ID NO:1.According to the type B acian metapneumovirus subunit vaccine, genetic engineering pichia pastoris X33-F for expressing the type B acian metapneumovirus F protein is inoculated into a culture medium, the type B acian metapneumovirus F protein is efficiently expressed under the action of methyl alcohol, the type B acian metapneumovirus F protein is extracted and purified and then added into a formaldehyde solution to be inactivated, then an oil adjuvant is added for mixing emulsification, and the vaccine is prepared.The prepared vaccine can increase the level of an immunized antibody and improve the uniformity of the immunized antibody, and the immune effect of the vaccine can be ensured; 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 a type B avian metapneumovirus subunit vaccine. Background technique [0002] Avian metapneumovirus belongs to the paramyxovirus pneumovirus genus, and its genome is a single-stranded negative-sense RNA without segments, which can cause respiratory diseases in turkeys, called turkey rhinotracheitis; after infecting chickens, it can cause viral air sacculitis, Rhinotracheitis and swollen head syndrome, decreased egg production. In the late 1970s, the disease caused by avian metapneumovirus was first reported in South Africa, and subsequently occurred in North America, South America, the Middle East, and the Far East. The virus was first isolated in the United States in 1989. While isolating the virus, it is often possible to isolate Bordetella, Pasteurella, Pseudomonas, Rhinotracheal Avian bacillus, and Alcaligenes faecalis. Avian metapneumovi...

Claims

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Application Information

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IPC IPC(8): A61K39/155A61P31/14
CPCA61K39/12A61K2039/552C12N2760/18334
Inventor 刘新文孙健邹敏程增青徐保娟郭玉广
Owner YEBIO BIOENG OF QINGDAO
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