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Coxsackie virus A6 strain (WF057R) and applications thereof

A Coxsackie virus and strain technology, applied in the direction of application, virus, antiviral agent, etc., can solve problems such as shortage, development constraints of anti-CVA6 drugs and vaccines, lack of stable animal infection models, etc.

Inactive Publication Date: 2017-07-14
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 hand, foot and mouth disease 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. A suitable animal model for CVA6 infection

Method used

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  • Coxsackie virus A6 strain (WF057R) and applications thereof
  • Coxsackie virus A6 strain (WF057R) and applications thereof
  • Coxsackie virus A6 strain (WF057R) and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1, Isolation and Identification of Coxsackievirus Type A6 (CVA6) Strain WF057R

[0058] In 2015, the inventor isolated a Coxsackievirus type A6 (Coxsackievirus) strain from the feces of a hand-foot-mouth child in Shandong Province, and named the strain WF057R. Coxsackievirus type A6 (Coxsackievirus) strain WF057R has been preserved in the General Microbiology Center of China Committee for the Collection of Microbial Cultures (CGMCC for short) on December 26, 2016. The address is: Courtyard 3, No. 1 Beichen West Road, Chaoyang District, Beijing No.), the deposit number is CGMCC No.13393.

Embodiment 2

[0059] Example 2, Utilizing Coxsackie A6 Virus Strain WF057R to Prepare CVA6 Infected Animal Model

[0060] 1. Cultivation of WF057R

[0061] RD cells were inoculated in medium 1 (medium 1 is the liquid obtained by adding fetal bovine serum, penicillin and streptomycin to MEM maintenance solution, wherein the mass percentage concentration of fetal bovine serum is 10%, and the mass percentage of penicillin Concentration is 1%, the mass percent concentration of streptomycin is 1%), in temperature is 37 ℃, 5% concentration CO 2 cultured in a cell incubator to obtain RD cell culture medium. The Coxsackie A6 virus strain WF057R of Example 1 was inoculated on the RD cells in the RD cell culture medium, and when the cells had a cytopathic effect (Cytopathic effect, CPE) area exceeding 80%, the cell culture medium was collected to obtain CVA6 strain WF057R virus liquid, quantify virus by limiting dilution method in 96-well plate, and measure virus TCID 50 Afterwards, it was aliquot...

Embodiment 3

[0093] Example 3. The antiserum produced by the active CVA6 vaccine using inactivated WF057R can protect mice and treat diseases caused by CVA6

[0094] 1. Preparation of inactivated CVA6 vaccine

[0095] The diluted CVA6 strain WF057R virus liquid (10 5 TCID 50 / ml) was diluted with formalin at a ratio of 1:4000 (V / V) to obtain a virus dilution, and the virus dilution was incubated at 37°C for 72 hours to obtain an inactivated virus solution; the inactivated virus mixed with complete Freund's adjuvant in equal volumes to make a complete emulsion, which is the CVA6 vaccine, and the content of inactivated virus in the CVA6 vaccine is 5×10 4 TCID 50 / ml. The infectious titer of CVA6 vaccine was detected by micro titration method to observe its inactivation effect.

[0096] Virus micro titration method: 1×10 per well 4 Cell / 100 μ l culture fluid (the cell in this culture fluid is RD cell, and substratum is the substratum 1 of embodiment 2) RD cell is inoculated on 96 wells ...

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Abstract

The invention discloses a Coxsackie virus A6 strain (WF057R) and applications thereof. The preservation number of the Coxsackie virus A6 strain (WF057R) in China General Microbiological Culture Collection Center (CGMCC) is CGMCC No.13393. A vaccine and CVA6 antiserum prepared from WF057R can treat and prevent diseases caused by CVA6. Furthermore, the maternal-transferred antibody of WF057R also has a protective effect on newborn mice. WF057R can also be used to establish a stable Coxsackie virus A6 infected animal model with good repeatability, and the animal model can be applied to drug antivirus treatment and immune protective effect evaluation of viral inactivation vaccines.

Description

technical field [0001] The invention relates to the Coxsackievirus A6 strain WF057R and its application in the field of biotechnology. Background technique [0002] Coxsackievirus (CV) is a single-stranded positive-sense RNA virus belonging to the family Picornaviridae and the genus Enterovirus. Coxsackievirus particles are icosahedral stereosymmetric, spherical in shape, about 23-33nm in diameter, nucleocapsid exposed, without envelope and protrusions; Coxsackievirus genome contains only one open reading frame (ORF), encoding a relatively long polyprotein precursors. The ORF is sequentially divided into three regions, P1, P2 and P3, wherein the P1 region encodes the structural proteins VP1-VP4 of the virus, which constitute the viral capsid. The P2 and P3 regions encode nonstructural proteins. [0003] Coxsackie virus is the main pathogen that causes hand, foot and mouth disease (HFMD) in children, especially infants under 3 years old, and can lead to severe neurological...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00C12N15/41C07K16/10C07K16/06A61K39/125A61K35/76A61K48/00A61K31/7088A61P31/14
CPCA61K35/76A61K39/12A61K48/005C07K16/06C07K16/1009C12N7/00C12N2770/32034C12N2770/32321
Inventor 史卫峰张振杰董兆鹏
Owner TAISHAN MEDICAL UNIV
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