狂犬病小鼠模型的制作

目的建立狂犬病病毒感染动物疾病模型。方法狂犬病病毒CVS-B2C毒株以10LD50剂量腿部肌肉注射接种4~6周龄的BALB/c小鼠,0.2 mL/只,于BSL2实验室负压IVC设备内饲养观察,并对其模型进行评价。结果小鼠接种狂犬病病毒后一周左右就出现临床症状,表现为饮食量下降,毛皮慢慢失去原先的光泽,体重下降,并出现麻痹等症状,进而死亡,部分小鼠出现躁狂的症状或抽搐性和强直性痉挛,而对照组小鼠则表现正常。DFA法检测结果：感染上狂犬病毒的小鼠脑组织涂片中出现特异性荧光抗体染色反应,而对照组动物的小鼠脑组织涂片未出现荧光抗体染色反应。RT-PCR法检测结果：从感染小鼠脑组织标本中提取病毒RNA,引物扩增出的目的基因片段,大小约为1kb,为N蛋白。免疫组化法检测结果：感染狂犬病毒的小鼠脑切片显示出棕色阳性颗粒,对照组小鼠脑切片染色阴性。病理检测结果：HE染色可见感染小鼠脑组织炎性细胞浸润,神经细胞胞质内出现内基体以及神经细胞退行性病变。结论成功的复制出小鼠狂犬病病毒感染模型,为研究和控制狂犬病奠定了基础。

A Method to Establish a Mouse Model of Rabies

Objective To establish an animal model of rabies. Methods Rabies virus strain CVS-B2C was inoculated into the leg i.m. in 4 - 6 week-old BALB/c mice in a dose of 10 LDSO. Then the mice were fed and observed in negative pressure IVC cages which located in BSL-2 laboratory. The mice were kifled and brain samples were taken for histopathological examination, immunohistochemistry, RT-PCR, DFA examination and rapid fluorescent focus inhibition test （RFFIT）. Results Clinical symptoms appeared about one week after inoculation, including decreased appetite, loss of fur luster, loss of body weight, and coma even death. Some mice appeared irritable, or showing twitch and entasia. The control mice appeared normal. The DFA examination showed specific fluorescence of anti-rabies N antibody in brain smears of the rabies virus-infected mice, but nothing in the control mice. RT-PCR revealed the virus RNA isolated from brain samples of the infected mice, and the size of the aim gene segment was 1 kb, which was rabies-N. The immunohistochemistry showed positive staining in the brain tissue of infected mice, but negative in the control mice. In the HE stained histological sections, intracellular Negri＇ s body in neurons,, neuron degeneration and inflammatory ceils could be seen in the brain tissue of infected mice. Conclusion A mouse model of rabies has been successfully established, which may be helpful in rabies research and control.