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Method for establishing high-efficiency virus infection model by centrifugation

A technology of virus infection and virus, which is applied in the field of virus infection, can solve the problems of low infection efficiency of virus to host cells, difficulty in separation and cultivation, etc., and achieve the effect of simple and easy method, improved infection efficiency, and increased infection rate

Inactive Publication Date: 2017-12-12
CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0006] In view of the above-mentioned shortcomings of the prior art, the object of the present invention is to provide a method for establishing a high-efficiency virus infection model using centrifugation, which is used to solve the problem of the low efficiency of virus infection of host cells in the prior art and the difficulty of removing from host cells. Issues such as isolation and culture that can advance the study of related viruses and the development of vaccines and antibodies against them

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  • Method for establishing high-efficiency virus infection model by centrifugation
  • Method for establishing high-efficiency virus infection model by centrifugation
  • Method for establishing high-efficiency virus infection model by centrifugation

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Embodiment 1

[0037] Materials and methods

[0038] Materials and reagents: Dulbecco's Modified Eagle Medium (DMEM) medium, blue-chain double antibody were purchased from Hyclone Company; trypsin was purchased from Sigma Company; fetal bovine serum was purchased from PAN Company; culture flasks and 24-well plates were purchased from Corning Company; Viral RNA extraction kit was purchased from Axygen.

[0039]Cell and virus culture: Vero E6 cell line was purchased from ATCC, and the cells were cultured in DMEM medium (cell culture medium) containing 10% (V / V) fetal bovine serum, 100 IU / mL penicillin and 100 μg / mL streptomycin . The virus medium is DMEM medium containing 3% fetal bovine serum (V / V), 100 IU / mL penicillin, 100 μg / mL streptomycin and 0.5 mg / mL trypsin. The virus used in the experiment was prepared by our research group using reverse genetic technology (see "Zhang Jinhui, Ge Jinying, Ren Yan, Bu Zhigao, Zhao Xiaodong for details of the preparation method. Using reverse genetic ...

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Abstract

The invention provides a method for establishing a high-efficiency virus infection model by centrifugation. The method comprises the following steps: inoculating a virus into cells, incubating for 15 minutes or more, centrifuging to obtain cells subjected to virus infection, wherein the virus is a virus entering the cells in an adhesion manner. According to a physical centrifugation method, the virus infection efficiency is improved, and the low-speed centrifugation method is simple, feasible and applicable to viruses entering the cells through adhesion.

Description

technical field [0001] The invention relates to the technical field of virus infection, in particular to a method for establishing a high-efficiency virus infection model by using centrifugation. Background technique [0002] Virus infection usually brings adverse symptoms such as diseases to humans or animals. Therefore, it is usually necessary to study viruses and their host cells, which then involves the establishment of in vitro virus infection models. Some viruses have poor susceptibility to host cells in vitro. , it is difficult to isolate and cultivate, and it is difficult to establish a corresponding infection model for research on the infection mechanism. [0003] For example, human metapneumovirus (human metapneumovirus, hMPV) is a respiratory virus newly isolated and discovered in 2001. It is a single negative-sense RNA virus and belongs to the genus Metapneumovirus of the subfamily Pneumoviridae of the family Paramyxoviridae. Since its discovery, there have been...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/86C12N5/10
CPCC12N15/86C12N5/0686C12N2510/00C12N2760/18343
Inventor 赵耀何浩陈素华杨晖
Owner CHILDRENS HOSPITAL OF CHONGQING MEDICAL UNIV