Chicken infectious bursal disease virus novel variant strain subunit vaccine and preparation method and application thereof

A subunit vaccine and bursal disease technology, applied in the fields of botany equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of antigenic vvIBDV differences, central immune organ damage, immune suppression effects, etc., to achieve The effect of highly effective immune protection

Active Publication Date: 2021-03-16
HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004]Our previous research found that the new mutant strain of IBDV not only caused central immune organ damage and immunosuppression, which significantly affected the prod

Method used

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  • Chicken infectious bursal disease virus novel variant strain subunit vaccine and preparation method and application thereof
  • Chicken infectious bursal disease virus novel variant strain subunit vaccine and preparation method and application thereof
  • Chicken infectious bursal disease virus novel variant strain subunit vaccine and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Isolation and identification of a new variant of chicken infectious bursal disease virus SHG19

[0055] 1. Materials and methods

[0056] 1.1 Clinical sample collection and processing

[0057] In September 2017, a 36-day-old broiler from a chicken farm in Chuzhou City, Anhui Province developed clinical symptoms of suspected IBD. The sick chickens showed lack of energy, and their bursa was found to be severely atrophied at autopsy. Add an appropriate amount of PBS to the sample of the diseased chicken bursa for inspection, grind and freeze-thaw three times. The prepared suspension is centrifuged at 4°C and 5000 g for 5 min, and the supernatant is taken for subsequent detection.

[0058] 1.2 SPF chicken, virus and main reagents

[0059] Specific-pathogen-free (SPF) chickens and SPF chicken embryos were provided by the Experimental Animal Center of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, and SPF chickens were raised in n...

Embodiment 2

[0117] Example 2 Preparation of IBDV novel variant virus-like particles

[0118] 1. Materials and methods

[0119] 1.1 Viruses, plasmids and strains

[0120] The representative strain of IBDV novel variant strain SHG19 (Fan et al., 2019) was isolated and preserved in Example 1. The pCold-I prokaryotic expression plasmid is a TaKaRa product. Escherichia coli engineering strain DH5α and E.coli Transetta (DE3) are products of Beijing Quanshijin Biotechnology Co., Ltd.

[0121] 1.2 Main reagents

[0122] The M-MLV Reverse Transcription Kit is an Invitrogen product. RNAiso Plus and PrimerStar DNAPolymerase are TaKaRa products. Restriction endonucleases KpnI and EcoRI, PrestainedProtein Ladder are ThermoFisher Scientific products. The gel recovery kit is an Axygen product. The homologous same group kit is a product of Nanjing Novizan Biotechnology Co., Ltd. Sepharose 6 Fast Flow columns are GE products. Ammonium sulfate is a product of Sinopharm Chemical Reagent Co., Ltd. ...

Embodiment 3

[0166] Example 3 Preliminary evaluation of the immune effect of IBDV novel variant subunit vaccine (SHG19-VP2)

[0167] 1 Materials and methods

[0168] 1.1 Main reagents, consumables and instruments

[0169] Fetal bovine serum is Ausbian product; EDTA-trypsin digestion solution and penicillin-streptomycin double antibody are products of Harbin Guosheng Biotechnology Co., Ltd.; DMEM and 1640 medium are Sigma products; cell culture plates are NEST products; The instrument is a PE product.

[0170] 1.2 Cells and viruses

[0171] DF1 cells were preserved by the Innovation Team of Avian Immunosuppressive Diseases (hereinafter referred to as the laboratory) of Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences. The representative strain SHG19 of the new variant of IBDV was isolated and identified in Example 1, and the representative strain of IBDV hypervirulent HLJ0504 was isolated and identified by our laboratory. IBDV recombinant virus rGtVarVP2 (w...

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Abstract

The invention discloses a chicken infectious bursal disease virus novel variant strain subunit vaccine and preparation method and application thereof. According to the vaccine and the preparation andthe application thereof, aiming at prevalence of an IBDV novel variant strain, a gene expression cassette of main protective protein VP2 of an IBDV novel variant strain representative strain SHG19 isdesigned, an element facilitating correct conformation formation of the gene expression cassette is introduced, recombinant prokaryotic expression plasmids are constructed, recombinant escherichia coli genetically engineered bacteria are prepared, through exploration of expression conditions and purification conditions, an IBDV novel variant strain virus-like particle is efficiently prepared, thevirus-like particle and adjuvant are emulsified to prepare the vaccine, and the test results show that the vaccine provides 100% immune protection for the homologous novel variant strain, and providesgood immune protection for the lethal attack of heterologous vvIBDV. The invention provides an effective technical means for the prevention and treatment of the the IBDV novel variant stain and a supervirulent strain.

Description

technical field [0001] The invention relates to a subunit vaccine of a new variant strain of IBDV, and a preparation method and application thereof, in particular to a subunit vaccine composed of VP2 protein virus-like particles of a new variant of IBDV Unit vaccine, its preparation method and application. The invention belongs to the technical field of veterinary medicine. Background technique [0002] Infectious bursal disease (IBD) is an important immunosuppressive disease of chickens, and its pathogen is infectious bursal disease virus (IBDV). The harm of IBDV to poultry industry is divided into direct harm and indirect harm. Direct harm: Destroy the central immune organ, the bursa, and can directly cause death. Indirect harm: IBDV infection-resistant chickens have low immunity, resulting in the failure of vaccine immunity against other important diseases such as avian influenza and Newcastle disease, and easy to cause secondary infection, affecting the production per...

Claims

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Application Information

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IPC IPC(8): C07K14/08A61K39/12A61P31/14C12N15/70C12N15/40
CPCC07K14/005C12N15/70A61K39/12A61P31/14C12N2720/10022C12N2720/10034C12N2720/10051C12N2720/10023Y02A50/30
Inventor 祁小乐王笑梅高玉龙高立李凯崔红玉刘长军潘青张艳萍刘爱晶
Owner HARBIN VETERINARY RES INST CHINESE ACADEMY OF AGRI SCI
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