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Coxsackievirus B5 CV-B5 and application thereof in preparing infectious animal models and kits

A technology for coxsackie virus, infecting animals, applied in the field of virology, can solve the problem that the animal model of brain lesions or spinal neuropathy has not been established effectively.

Pending Publication Date: 2017-10-10
NAT INST FOR FOOD & DRUG CONTROL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] It can be seen that although CV-B5 virus is one of the main pathogens of aseptic meningitis, it has not yet been possible to establish an animal model for effectively obtaining brain lesions or spinal cord neuropathy.

Method used

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  • Coxsackievirus B5 CV-B5 and application thereof in preparing infectious animal models and kits
  • Coxsackievirus B5 CV-B5 and application thereof in preparing infectious animal models and kits
  • Coxsackievirus B5 CV-B5 and application thereof in preparing infectious animal models and kits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1 Isolation and establishment of CVB5 / JS417 virus strain

[0033] Taking the clinical samples of hand, foot and mouth cases in Jiangsu area collected in 2013 as the research object, a virus strain that can replicate stably was obtained by separating and purifying VERO cells, named CVB5 / JS417, and preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee , the deposit number is CGMCC No.13851. After the amplification of the whole gene sequence, the comparison of the gene sequence confirmed that it is a Coxsackievirus B Group 5 type virus (Coxsackievirus B Group 5type), and the full length of the gene is 7370bp. The detection of exogenous factors confirmed that the virus strain was not contaminated by other exogenous factors. According to the analysis of biological characteristics, the virus titer of the virus strain is 8.50LgCCID50 / ml.

[0034] Among them, sampling of pathogens, cell separation and purificatio...

Embodiment 2

[0035] The establishment of embodiment 2 CV-B5 animal model

[0036] (1) Selection of animal strains

[0037]BALB / c, C57, NIH, ICR, and Kunming mice aged 1 to 3 days were selected for intraperitoneal infection with the same dose of CVB5 / JS417 virus. After 3 days, the experimental animals successively showed hindlimb paralysis, malaise and even death. Among them, 70% to 100% of the experimental mice in the BALB / c and C57 mouse groups died after 6 days of infection.

[0038] Considering that BALB / c mice are inbred mice with a more similar genetic background, which is conducive to the consistency of the experiment, BALB / c mice were selected to establish the CV-B5 animal model for further experiments.

[0039] (2) Selection of the age of suckling mice

[0040] BALB / c mice of 1, 3, 5, 7, and 14 days old were intraperitoneally challenged with the same dose of CVB5 / JS417 virus strain, and the survival rate was observed.

[0041] Such as figure 2 As shown, CVB5 / JS417 can cause mo...

Embodiment 3

[0052] Embodiment 3 pathological conditions

[0053] Set up the CV-B5 mouse experimental animal model according to the mode described in Example 2 section (4), collect respectively the brain, heart, liver, kidney, lung, spleen, spinal cord and other tissues of dying experimental mice after the attack for pathological examination and Immunohistochemical staining.

[0054] The control group was obtained with physiological saline according to the same test method, collection time and inspection method as the experimental animals.

[0055] The pathological examination adopts tissue fixation, paraffin section, pathological HE staining and immunohistochemical staining, and microscopic observation, and judges the results based on tissue sections of normal newborn mice and negative control mice. The judgment of pathological examination and immunohistochemical staining results are routine means in the field.

[0056] Histopathological results such as Figure 6 As shown, obvious eosi...

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Abstract

The invention relates to Coxsackievirus B5 CV-B5 CVB5 / JS417 capable of being stably copied and abdicated. The preservation number of the virus is CGMCC (China general microbiological culture collection center) No. 13851. The invention further relates to an application of Coxsackievirus B5 CV-B5 the CVB5 / JS417 in preparing a Coxsackievirus B5 CV-B5 infected mouse. The experimental results show that obvious eosinophilic necrosis and vacuolar degeneration necrosis in spots of the brain, spinal cord, cardiac muscle and skeletal muscle of hind limbs of the Coxsackievirus B5 CV-B5 infected mouse can be seen, and meanwhile, immunohistochemistry shows that positive or strong positive reaction appears in brain and spinal neural cells and cardiac muscle and skeletal muscle cells of the hind limbs, which verifies that the CVB5 / JS417 can cause obvious lesions and necrosis to the brain, spinal nerves, cardiac muscle and skeletal muscle of the hind limbs of the infected mouse.

Description

【Technical field】 [0001] The present invention relates to virology. In particular, the present invention relates to a strain of Coxsackie B group 5 virus capable of infecting suckling mice, and the use of the virus. 【Background technique】 [0002] Coxsackievirus B5 type (Coxsackievirus B5, CV-B5) belongs to Picornaviridae (Picornaviridae), Enterovirus (Enterovirus). CV-B5 is one of the main pathogens of aseptic meningitis. In addition to upper respiratory tract infection, diarrhea and hand, foot and mouth disease, CV-B5 infection mainly causes viral encephalitis, aseptic meningitis, pancreatitis, flaccid paralysis, dilated myocarditis and diabetes and other severe diseases. In addition, according to literature reports, CV-B5 infection can cause rare clinical symptoms such as transient aphasia and acute transverse myelitis in children. In recent years, a high rate of CV-B5 infection has been detected in HFMD cases in my country, but there are few related basic studies. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A01K67/027A61K39/125A61P31/14C12R1/93
CPCA01K67/0271A01K2207/20A01K2227/105A01K2267/0337A61K39/12A61K2039/5252C12N7/00C12N2770/32321C12N2770/32334
Inventor 毛群颖郝晓甜梁争论李秀灵
Owner NAT INST FOR FOOD & DRUG CONTROL
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