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Method for building 7-day-old mouse model infected with enterovirus 71

A technology of mouse model and enterovirus, which is applied in the establishment of 7-day-old mouse model, can solve problems such as model instability, EV71 pathogenic mechanism, and difficulty in developing clinical vaccines, achieving broad application prospects

Inactive Publication Date: 2011-03-02
INST OF BASIC MEDICINE OF SAMS
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  • Summary
  • Abstract
  • Description
  • Claims
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Problems solved by technology

At present, the animal models used at home and abroad are suckling mice within 4 days of age, but this model is very unstable, which brings great difficulties to the pathogenic mechanism, clinical treatment and vaccine development of EV71

Method used

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  • Method for building 7-day-old mouse model infected with enterovirus 71
  • Method for building 7-day-old mouse model infected with enterovirus 71
  • Method for building 7-day-old mouse model infected with enterovirus 71

Examples

Experimental program
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Effect test

Embodiment 1

[0015] Embodiment 1 establishes the mouse model infected with enterovirus 71

[0016] Proceed as follows:

[0017] 1. Isolation and identification of EV71

[0018] 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 patients ...

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Abstract

The invention discloses a method for building a 7-day-old mouse model infected with enterovirus (EV) 71, comprising the following steps: taking 7-day-old suckling mice and injecting liquid virus into the brains of the mice, thus obtaining the mouse model infected with EV 71. The injection volume of the liquid virus is 20mu L and the liquid virus is prepared by the following steps: inoculating the EV71 seed into MA104 cells in terms of inoculation amount of 200mu L / 100mL cell culture bottle, culturing the MA104 cells in 5% of CO2 at 37 DEG C and observing the cytopathic effect every day; on the second day after inoculating EV71, observing that about 70% of the MA104 cells are agglutinated and crimpled, freeze-thawing the cells in the whole bottle and then centrifuging the cells and taking the supernatant, thus obtaining the liquid EV71. The method successfully builds the stable 7-day-old mouse model infected with EV71, provides foundations for the scientific research personnel to research the EV71 etiology and pathogenic mechanism and for screening efficient antivirus drugs and developing vaccines and has broad application prospect.

Description

technical field [0001] The invention relates to a method for establishing a 7-day-old 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 causing hand, foot and mouth disease in infants and young children. 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. A...

Claims

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Application Information

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IPC IPC(8): A61K35/76
Inventor 李鹏岳盈盈宋楠楠李志会孟红
Owner INST OF BASIC MEDICINE OF SAMS
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