Vaccine for preventing and treating diseases caused by Coxsackie virus and preparation method and application thereof

A coxsackie virus and vaccine technology, applied in chemical instruments and methods, antiviral immunoglobulins, biochemical equipment and methods, etc., can solve the development constraints of anti-CVA6 drugs and vaccines, and the lack of stable animals for anti-CVA6 drugs and vaccines. Infection model, unclear pathogenesis of hand, foot and mouth disease, etc.

Inactive Publication Date: 2017-08-01
TAISHAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the pathogenesis of HFMD is still unclear, especially the pathogenic mechanism of CVA6 has not been reported, and the screening of anti-CVA6 drugs and vaccines lacks stable animal infection models, and the development of anti-CVA6 drugs and vaccines is also restricted

Method used

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  • Vaccine for preventing and treating diseases caused by Coxsackie virus and preparation method and application thereof
  • Vaccine for preventing and treating diseases caused by Coxsackie virus and preparation method and application thereof
  • Vaccine for preventing and treating diseases caused by Coxsackie virus and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1. Isolation and identification of Coxsackie virus A6 (CVA6) strain WF057R

[0056] The inventors isolated a Coxsackievirus A6 (Coxsackievirus) strain from the feces of a child with hand, foot and mouth in Shandong Province in 2015, and named the strain WF057R. Coxsackievirus A6 (Coxsackievirus) strain WF057R was deposited on December 26, 2016 in the General Microbiology Center of the China Microbial Culture Collection Management Committee (abbreviated as CGMCC, address: No. 3, Beichen West Road, Chaoyang District, Beijing) No.), the deposit number is CGMCC No.13393.

Embodiment 2

[0057] Example 2: Preparation of CVA6 infection animal model using Coxsackie A6 virus strain WF057R

[0058] 1. Cultivation of WF057R

[0059] Inoculate RD cells in medium 1 (medium 1 is the liquid obtained by adding fetal bovine serum, penicillin and streptomycin to the MEM maintenance solution, wherein the mass percentage of fetal calf serum is 10%, and the mass percentage of penicillin The concentration is 1%, the mass percentage concentration of streptomycin is 1%), the temperature is 37℃, the concentration of 5% CO 2 Cultivate in a cell incubator to obtain RD cell culture solution. The Coxsackie A6 virus strain WF057R of Example 1 was inoculated on the RD cells in the RD cell culture medium, and when the area of ​​the cells with cytopathic effect (CPE) exceeded 80%, the cell culture medium was collected to obtain CVA6 strain WF057R virus solution, quantify the virus in a 96-well plate by limiting dilution method, and determine the virus TCID 50 Afterwards, it is divided into ...

Embodiment 3

[0092] Example 3. The antiserum produced by the CVA6 vaccine with inactivated WF057R as the activity can protect mice and treat diseases caused by CVA6

[0093] 1. Preparation of inactivated CVA6 vaccine

[0094] The diluted CVA6 strain WF057R virus liquid (10 5 TCID 50 / ml) Dilute with formalin at a ratio of 1:4000 (V / V) to obtain a virus dilution. Incubate the virus dilution at 37°C to inactivate the virus for 72 hours to obtain an inactivated virus solution; Mix the same volume with complete Freund’s adjuvant to make a complete emulsion, which is the CVA6 vaccine. The content of inactivated virus in the CVA6 vaccine is 5×10 4 TCID 50 / ml. The infectivity titer of CVA6 vaccine was detected by microtitration to observe its inactivation effect.

[0095] Virus microtitration method: press 1×10 per well 4 Cells / 100μl culture medium (the cells in the culture medium are RD cells, and the medium is the medium 1 of Example 2). Inoculate RD cells on a 96-well flat-bottom cell culture plate...

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Abstract

The invention discloses a vaccine for preventing and treating diseases caused by Coxsackie virus and a preparation method and application thereof. The preparation method of the vaccine for preventing and treating the diseases caused by Coxsackie virus comprises the following steps: taking an inactivated Coxsackie virus A6 strain as an active ingredient to prepare the vaccine; preservation number of the Coxsackie virus A6 strain in a general microorganism center of China Committee for Culture Collection of Microorganisms is No. CGMCC No. 13393. The vaccine and CVA6 antiserum prepared by CVA6 strain WF057R have the functions of treating and preventing the diseases caused by CVA6, and a parent antibody of CVA6 strain WF057R has protective effect on neonatal suckling mice.

Description

Technical field [0001] The invention relates to a vaccine for preventing and treating diseases caused by Coxsackie virus in the field of biotechnology, and a preparation method and application thereof. Background technique [0002] Coxsackievirus (CV) is a single-stranded positive-stranded RNA virus, belonging to the Picornaviridae family and enterovirus genus. Coxsackie virus particles are icosahedral three-dimensionally symmetrical, spherical, with a diameter of about 23-33nm, with naked nucleocapsid, no envelope and protrusions; the Coxsackie virus genome contains only one open reading frame (ORF), which encodes a relatively large Long polyprotein precursor. The ORF is divided into 3 regions, P1, P2, and P3, in which the P1 region encodes the structural proteins VP1 to VP4 of the virus, which constitute the viral capsid. The P2 and P3 regions encode non-structural proteins. [0003] Coxsackie virus is the main pathogen causing hand-foot-mouth disease (HFMD) in children, espec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/125A61P31/14C12N7/00C07K16/10C07K16/06
CPCA61K39/12A61K2039/545C07K16/06C07K16/1009C12N7/00C12N2770/32321C12N2770/32334
Inventor 史卫峰张振杰董兆鹏
Owner TAISHAN MEDICAL UNIV
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