人季节性H1N1流感病毒小鼠感染模型的建立

目的制备人季节性H1N1流感病毒的小鼠感染模型,为研究流感病毒致病性、研发抗病毒药物提供模型动物。方法将人季节性H1N1流感病毒在鸡胚尿囊腔扩增后,滴鼻接种小鼠,4 d后将小鼠处死,挑选感染体征严重者进行实时荧光PCR(FQ-PCR)检测肺中的流感病毒,将检测阳性的肺上清在鸡胚尿囊腔扩增,接种于下一代小鼠。比较各代小鼠对流感病毒的适应情况,直至小鼠出现明显的感染体征,取肺研磨制成匀浆,获得流感病毒鼠肺适应株并检测其半数致死量(LD50)。将10 LD50的病毒液接种于小鼠,建立人季节性H1N1流感病毒的小鼠感染模型,观察模型小鼠的一般活动状态、体质量变化、肺部病变,HE染色观察肺部病理切片,计算肺指数,FQ-PCR检测病毒RNA。结果人季节性流感病毒在小鼠体内传代4次后,鼠肺适应株制备完成,其经鼻的LD50为10-2.41/0.05 mL。人季节性流感病毒的小鼠感染模型,一般状态差,体质量明显减轻,肺指数增大,70%出现死亡。病理切片观察病变明显,FQ-PCR显示流感病毒阳性。结论成功建立了人季节性H1N1流感病毒的小鼠感染模型。

Establishment of a mouse infection model of human seasonal H1N1 influenza virus

Objective To establish a mouse infection model of human seasonal influenza virus(H1N1),and make preparations for the study on influenza viral pathogenicity and new antiviral drugs.Methods Mice were inoculated with influenza virus(H1N1) by the nasal cavity.After 4 days,mice with serious infectious signs were killed.A pneumonic homogenate that was confirmed with influenza virus infection by real time fluorescence quantitative polymerase chain reaction(FQ-PCR) was injected into the chick embryo allantoic cavity.Allantoic fluid was inoculated in to the next generation of mice.When infected mice presented obvious infectious signs,the 50% lethal dose(LD50) was calculated.The virus of 10LD50 was inoculated into mice to establish the infection model.Then,general condition,weight,lung index,and pathological sections were observed and viral RNA was detected by FQ-PCR.Results After 4 times of passage,the mouse infection model of human seasonal influenza virus(H1N1) was established,with LD50 of 10-2.41/0.05 mL.The mouse infection model had serious infectious signs,poor general condition,decreased weight, increased lung index,positive results by FQ-PCR and mortality of 70%.Conclusion The mouse infection model of human seasonal influenza virus(H1N1) has been successfully established.