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Raccoon dog parvovirus virus-like particle, preparation method thereof and application thereof as well as vaccine prepared by virus-like particle

A parvovirus, virus-like technology, applied in botanical equipment and methods, biochemical equipment and methods, applications, etc., can solve the problems of low immune efficacy and strong virulence of live vaccines, and achieve good immunogenicity. Immunity effect and preventive effect, low production cost effect

Active Publication Date: 2018-11-13
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Aiming at the problem that the current raccoon dog enteritis inactivated vaccine has low immune efficacy and the live vaccine has strong virulence, the invention provides a kind of raccoon parvovirus virus-like particle, and the virus-like particle is composed of a recombinant protein expressing raccoon parvovirus RDPV-VP2 protein. SF21 cells were transfected with baculovirus and cultured until the cells were infected

Method used

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  • Raccoon dog parvovirus virus-like particle, preparation method thereof and application thereof as well as vaccine prepared by virus-like particle
  • Raccoon dog parvovirus virus-like particle, preparation method thereof and application thereof as well as vaccine prepared by virus-like particle
  • Raccoon dog parvovirus virus-like particle, preparation method thereof and application thereof as well as vaccine prepared by virus-like particle

Examples

Experimental program
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Effect test

Embodiment 1

[0033] Example 1. Preparation method of raccoon parvovirus virus-like particles.

[0034] (1) Extraction of viral genome:

[0035] Isolation of raccoon parvovirus RDPV-HeB17 strain: Take the intestinal contents of dead raccoon dogs with enteritis, dilute the contents 10 times with 0.2M PBS, inoculate them on F81 cells according to the MOI of 0.1-0.3, and change the contents after the cells adhere to the wall. MEM medium with 2% newborn bovine serum was cultured in a carbon dioxide incubator at 37°C, and the lesions were observed every day. When 70% of the cells showed specific lesions such as cell pulling and shedding, they were harvested and frozen, and frozen and thawed twice at -20°C.

[0036] The isolated raccoon parvovirus RDPV-HeB17 strain was simultaneously inoculated into F81 cells, cultured at 37°C for 3-5 days, and after more than 75% of the lesions were detected, it was frozen and thawed twice, and DNA was extracted from the harvested virus liquid. For specific step...

Embodiment 2

[0131] Example 2. Application of raccoon parvovirus virus-like particles in the preparation of raccoon parvovirus enteritis vaccine:

[0132] 1. Identification of raccoon parvovirus virus-like particles:

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Abstract

The invention discloses a raccoon dog parvovirus virus-like particle, a preparation method thereof and application thereof as well as a vaccine prepared by the virus-like particle, and belongs to thefield of recombinant vaccine preparation. In order to solve the problem that raccoon dog parvovirus enteritis inactivated vaccine is relatively low in immunity and virulence of the live vaccine is strong, the invention provides the raccoon dog parvovirus virus-like particle. A raccoon dog parvovirus gene RDPV-VP2-OPTI optimized through a codon is connected with an insect expression vector pFastBac1 to obtain a donor plasmid; the donor plasmid is converted into baculovirus DH10 Bac E.coli to culture to obtain recombinant baculovirus for expressing raccoon dog parvovirus RDPV-VP2 protein, is transfected to an SF21 cell, and is cultured until the cell is infected to collect a cell virus solution. After the raccoon dog parvovirus virus-like particle is mixed with an aluminum hydroxide gel adjuvant, the raccoon dog parvovirus enteritis vaccine can be obtained. The raccoon dog parvovirus virus-like particle is suitable for preparing the raccoon dog parvovirus enteritis vaccine.

Description

technical field [0001] The invention belongs to the field of preparation of recombinant vaccines, and in particular relates to a raccoon parvovirus virus-like particle, its preparation method and application, and a vaccine prepared by the virus-like particle. Background technique [0002] Raccoon parvovirus enteritis mainly causes raccoon dog enteritis, and the mortality rate can reach 30%-60%. At present, the main means of preventing the disease is vaccine immunization. The existing vaccines are mainly inactivated whole virus vaccines or live vaccines. The main disadvantage is that the inactivated vaccine has low immune efficacy, and the live vaccine has the risk of strong virulence. Contents of the invention [0003] Aiming at the problem that the current raccoon dog enteritis inactivated vaccine has low immune efficacy and the live vaccine has strong virulence, the invention provides a kind of raccoon parvovirus virus-like particle, and the virus-like particle is compos...

Claims

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Application Information

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IPC IPC(8): C07K14/015C12N15/866A61K39/23A61P31/20
CPCA61K39/12A61K2039/5258A61K2039/55505A61P31/20C07K14/005C12N15/86C12N2710/14043C12N2750/14023C12N2750/14034
Inventor 罗国良王振军程悦宁易立冯二凯郭利程世鹏
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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