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Method for establishing enterovirus 71 (EV71) infected C57/BL6 suckling mouse model

An enterovirus and the technology of establishing a method, applied in the direction of medical materials derived from viruses/bacteriophages, etc., can solve the problems of exacerbation of infection, lack of therapeutic drugs and vaccines, EV71 infection route, pathogenic mechanism and epidemic law are not clear, etc., to achieve The effect of broad application prospects

Inactive Publication Date: 2012-03-21
济南市传染病医院
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Problems solved by technology

Since the outbreak of hand, foot and mouth disease in Fuyang, Anhui in 2008, the infection of EV71 has been intensified and has become a public health concern. Therapeutics and Vaccines

Method used

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  • Method for establishing enterovirus 71 (EV71) infected C57/BL6 suckling mouse model
  • Method for establishing enterovirus 71 (EV71) infected C57/BL6 suckling mouse model

Examples

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Embodiment 1

[0013] Example 1 Establishment of a suckling mouse model infected with enterovirus 71

[0014] Proceed as follows:

[0015] 1. Isolation and identification of EV71

[0016] The EV71 strain used in the present invention is isolated from the stool specimens of clinical severe hand-foot-mouth patients, and simultaneously uses human rhabdomyosarcoma cell line (RD), rhesus monkey kidney cell line (MA104) and African green monkey kidney cell line (Vero) Three kinds of cells were used to separate the viruses in the specimen. As a result, the bacteria on the three kinds of cells showed cytopathic changes, and the viruses were frozen and preserved after passage. Then extract the genomic RNA of the isolated EV71, reverse transcribe into cDNA, adopt the EV71 national standard primer to identify the isolated strain, the results show that the isolated strain has a very bright band at 226bp (such as figure 1 Shown), it can be confirmed that the strain isolated from the stool specimens of ...

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Abstract

The invention discloses a method for establishing an enterovirus 71 (EV71) infected C57 / BL6 suckling mouse model, which comprises the following step of: performing intramuscular injection on the hind legs of a C57 / BL6 suckling mouse to obtain the EV71 infected mouse model, wherein 20mu L of virus solution is injected. A method for preparing the virus solution comprising the following steps of: inoculating 200mu L of EV71 virus seed into MA104 cells in 100mL of cell culture flask, culturing at the temperature of 37 DEG C under the condition of 5 percent CO2, observing the condition of a cytopathic effect every day, freezing and thawing the whole flask of cells when 70 percent of MA104 cells are agglutinated and shrunk, and centrifuging to obtain supernatant, namely the EV71 virus solution. A stable EV71 injected C57 / BL6 suckling mouse model is successfully established, a basis is provided for studying the etiology and pathogenic mechanism of EV71, screening high-efficiency antiviral medicines and preparing vaccines, and the method is good in application prospect.

Description

technical field [0001] The invention relates to a method for establishing a C57 / BL6 suckling mouse model infected with enterovirus 71. Background technique [0002] Enterovirus 71 (EV71) belongs to the Enterovirus genus of the Picornaviridae family and is one of the main pathogens that cause Hand Foot and Mouth Disease (HFMD) in infants. The main symptoms of EV71 infection are rashes or herpes on the hands, feet, mouth and other parts. Some children may develop neurological complications such as aseptic meningitis, brainstem encephalitis, neurogenic pulmonary edema, and polio-like paralysis. Respiratory tract infection and myocarditis etc., can be disabled, lethal. Since the outbreak of hand, foot and mouth disease in Fuyang, Anhui in 2008, the infection of EV71 has been intensified and has become a public health concern. Therapeutic drugs and vaccines. [0003] The lack of mature and stable animal models is one of the most important reasons for the above situation. At p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/76
Inventor 张颖盖中涛
Owner 济南市传染病医院
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