Novel EV71 virus strain and application thereof

A new type of viral technology, applied in the direction of antiviral agents, viruses/phages, medical raw materials derived from viruses/phages, etc., can solve the problem of inability to accurately evaluate the therapeutic effect of immunogenic HFMD vaccines, weak therapeutic effects, etc. question

Active Publication Date: 2013-10-30
SOUTH CHINA UNITED VACCINE INST
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

However, the mortality rate of the treatment group still reached 70%, indicating that the in vivo treatment effect is weak
[0034] At present, there are no effective animal infe

Method used

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  • Novel EV71 virus strain and application thereof
  • Novel EV71 virus strain and application thereof
  • Novel EV71 virus strain and application thereof

Examples

Experimental program
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Embodiment Construction

[0041] Isolation and screening of virus strains:

[0042] Vero cell culture method

[0043] (1) Vero cell resuscitation (ATCC code: CCL-81): Take out a frozen cell from liquid nitrogen and quickly melt it in warm water at 39°C, then press the cell suspension to about 1.0×10 6 Inoculation / ml transfer to 175cm 2 In the square flask, gradually add medium (M199 medium containing 10% fetal bovine serum (vol / vol) (see GIBCO medium manual for the components of M199 dry powder medium)) to 60ml, 37℃, 5% CO 2 Cultivate in an incubator.

[0044] (2) Prepare a 24-well plate: culture the cells of (1) for 72 hours, digest with 0.25% trypsin, add 10% serum M199 medium to stop the reaction, add medium after counting to make the cell suspension density 1.0 ~2.0×10 5 Pcs / ml, pipette to disperse evenly, then add 1ml to each well of the 24-well plate, 37℃, 5% CO 2 Incubate in an incubator for 48 hours.

[0045] (3) Virus isolation, cultivation, and preservation

[0046] Retrieve clinical samples from the ...

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Abstract

The invention discloses a novel EV71 virus strain of which the whole-genome length is 7405bp and the sequence is shown in SEQID NO:1. The virus strain can infect ICR, KM, BALB/c and NIH mice; the virus dosage of larger than 1*10<2>-1*10<7> TCID50 can cause the disease attack or death of 1-17 days mice; the administration route can be intraperitoneal injection, encephalocoele injection and intragastric administration; the symptoms are typical symptoms of EV71 virus infection and are divided into the following five levels according to stages of the disease: Level 0: not attacked; L1: motion incoordination and weakness of limbs; L2: hypokinesia and paralysis of fore or posterior limbs; L3: quadriplegia and dying; and L4: death. Various animal evaluation models such as an immunogenicity animal evaluation model of hand-foot-and-mouth disease vaccine and an animal evaluation model of EV71 therapeutic drug and therapeutic vaccine, can be built conveniently.

Description

Technical field [0001] The invention relates to a new type of EV71 virus and its application. Background technique [0002] Epidemiology [0003] Hand, foot and mouth disease is a global infectious disease. The epidemic has been reported in most parts of the world. It was first described by Seddon of New Zealand in 1957. In 1974, Schmidt first isolated and identified EV71 from a stool sample of a child with a brain (1969). In the following 3 years, the virus was again isolated from 23 patients with severe nervous system in California. And then the disease has a broader spectrum of changes, with symptoms ranging from fever, hand-foot-mouth disease, herpes, aseptic meningitis, paralysis, encephalitis, and even death. [0004] In 1975, more than 750 people in Bulgaria became ill, and 44 of them died. [0005] In 1978, a large-scale epidemic occurred in Japan. A total of 36,301 cases were found. Except for a few aseptic meningitis, the main manifestation was HFMD. [0006] After 1997, th...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K39/125A61P31/14A61K35/76C12R1/93
Inventor 彭涛马书智廖鹏云王弋刘钿莲
Owner SOUTH CHINA UNITED VACCINE INST
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