Method for establishing enterovirus71 intramuscular injection-infected suckling mouse model
An intramuscular injection, enterovirus technology, applied in the direction of virus antigen components, pharmaceutical formulations, preparations for in vivo experiments, etc., can solve the problems of large animal damage, unfavorable animal models, abnormal animal death, etc., and achieve broad application prospects. Effect
Inactive Publication Date: 2012-11-28
INST OF BASIC MEDICINE OF SAMS +1
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Problems solved by technology
At present, most of the suckling mouse models used at home and abroad are infected with the virus through intracerebral injection, but this method of inoculation causes great damage to the animals and can easily lead to abnormal death of the animals, which is not conducive to the establishment of a stable animal model
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Embodiment 1
[0015] Example 1 Establishment of a suckling mouse model infected with enterovirus 71 by intramuscular injection
[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 f...
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The invention discloses a method for establishing an enterovirus71 (EV71) intramuscular injection-infected suckling mouse model. The EV71 intramuscular injection-infected suckling mouse model is obtained by performing intramuscular injection of virus solution on a BALB / c suckling mouse. The virus solution is prepared by the following steps of: inoculating the EV71 viruses into MA104 cells in an inoculum size that 200muL of viruses are inoculated in a 100mL cell culture bottle, culturing 5 percent CO2 at the temperature of 37 DEG C and observing the pathological change condition of the cells every day; wherein about 70 percent of MA104 cells are agglutinated and crimped on the second day after the inoculation of the EV71; and freeze-thawing the entire bottle of cells, centrifuging and taking the supernatant to obtain the virus solution of the EV71. By the method, a stable EV71 intramuscular injection-infected suckling mouse model is successfully established, a foundation is provided for the research personnel to research EV71 type etiology, EV71 type pathogenic mechanism and screening and vaccine research of high-efficiency anti-virus medicaments.
Description
technical field [0001] The invention relates to a method for establishing a suckling mouse model infected by intramuscular injection of enterovirus 71. Background technique [0002] Hand, foot and mouth disease (HFMD) is a global infectious disease caused by enterovirus, and there are reports of the prevalence of this disease in most parts of the world. Among them, HFMD caused by enterovirus type 71 (EV71) is the most serious, and complications such as myocarditis, pulmonary edema, meningoencephalitis, and polio-like paralysis often occur, and severe cases lead to death. EV71 belongs to the genus Enterovirus of the picornaviridae family, similar to the poliovirus of the same genus, EV71 has a certain tissue tropism to the anterior horn cells of the spinal cord, and is the most common non-polio enterovirus causing acute flaccid paralysis Therefore, EV71 is considered to be the most important neurotropic enterovirus after poliovirus. [0003] Due to the lack of animal models...
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IPC IPC(8): A61K49/00A61K39/12A61K35/765
Inventor 岳盈盈张颖李鹏李志会宋楠楠盖中涛孟红
Owner INST OF BASIC MEDICINE OF SAMS