EV-71(Enterovirus 71) type viral plaque purification method

A technology of 1. EV-71 and virus plaque, applied in the direction of recovery/purification, can solve problems such as health hazards of infants and young children, and achieve fast and accurate purification, good application prospects, and easy operation

Inactive Publication Date: 2011-01-19
MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
View PDF1 Cites 7 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the disease has been reported before, it mostly appeared as sporadic cases. In 2009, it broke out in many areas of Anhui, Henan and Hebei in my country, causing serious harm to the health of infants and young children.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • EV-71(Enterovirus 71) type viral plaque purification method
  • EV-71(Enterovirus 71) type viral plaque purification method
  • EV-71(Enterovirus 71) type viral plaque purification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1, the establishment of EV-71 plaque formation method

[0028] 1. Materials and methods

[0029] 1) Cells: Vero cell strain (purchased from Invitrogen), subcultured according to conventional methods, and monolayer cells grown within 48 hours were used for experiments.

[0030] 2) Sample to be purified:

[0031] Three clinical stool samples of HFMD patients were collected from Henan Province, one clinical throat swab sample of HFMD patients was collected from Anhui Province, and three clinical anal swab samples of HFMD patients were collected from Chongqing City. The stool samples were dissolved separately. In a sterile pH7.2, 0.01M / L PBS solution (containing 2.5ug / ml amphotericin B or 40ug / ml nystatin), mix well and centrifuge, and pass the supernatant through a 0.22um Inoculate Vero monolayer cells directly after the filter membrane; after the throat swab and anal swab samples are collected, break the cotton swab and put it into 2ml of bovine serum albumin...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses an EV-71(Enterovirus 71) type viral plaque purification method, comprising the following steps of: (1) preparing monolayer vero cell, and adding a sample solution containing virus for adsorption for 1 hour; (2) covering a layer of MEM (Minimum Essential Medium) solid culture medium on the cell subjected to adsorption, and culturing until a viral plaque appears; and (3) andpicking the viral plaque to obtain a viral solution. In the invention, by adsorbing a vero cell monolayer with a virus sample and covering with the MEM cell solid culture medium, obvious plaque can be observed in 3-5 days. Shown by experiments, after three rounds of plaque purification and amplification, a purified virus strain can be obtained on the basis of RT-PCR (Reverse Transcription-Polymerase Chain Reaction) detection and sequencing.

Description

technical field [0001] The invention relates to a method for plaque purification of EV-71 virus. Background technique [0002] At present, there are also reports on plaque purification technology. Firstly, inoculate the virus liquid with different dilutions in the culture vessel. After absorbing for 1-4 hours, the adsorbed virus liquid is discarded and the residual virus liquid is washed away with the maintenance solution. After the first nutrient covering layer is solidified, turn it over and incubate for several days, then add the second layer of covering layer. After solidification, turn it over and incubate in the dark for 1-2 days to observe the formation of plaques. However, this purification method may cause the virus to spread from the plaque to the agar surface and contaminate other plaques in the same culture. This is because when the virus is multiplied and then added to the neutral red solution or the second layer of agar, it will Viruses disperse on the agar su...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/02
Inventor 常国辉吴晓燕林磊罗彦军户义李靖杨银辉康晓平祝庆余
Owner MICROBE EPIDEMIC DISEASE INST OF PLA MILITARY MEDICAL ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap