建立H5N1流感病毒感染雪貂动物模型

Establish H5N1 Flu Virus Infection Ferret Animal Model

目的建立甲型H5N1流感病毒感染雪貂动物模型[1-3]。方法 H5N1流感病毒株A/Vietnam/1203/2004病毒以103和104 TCID50滴度分别感染雪貂。对4～10月龄去势雪貂经兽用氯胺酮轻度麻醉后进行滴鼻感染,每个稀释度接种3只雪貂,感染后第5天安乐处死。感染后每天记录雪貂一般临床变化。感染前0 d采集鼻甲骨活检,感染后1～5 d鼻甲骨活检检测病毒载量和病毒滴度。处死时取雪貂气管、肺、心、肝、脾、肾、小肠、脑组织作病毒滴度检测和病理检查。结果 H5N1103和104 TCID50的病毒分别感染雪貂,雪貂死亡都率在33%。103TCID50和104 TCID50病毒分别感染雪貂,动物都出现持续3 d体温升高,104 TCID50组出现超过20%的体重下降。上呼吸道排毒呈现上升趋势,并可在除呼吸系统以外的组织器官中分离到病毒。感染的雪貂病理表现为重度肺炎。结论雪貂感染H5N1病毒株后在临床表现、病毒学、分子生物学、病理学方面的检测都可以证实雪貂感染H5N1病毒动物模型已建立,104 TCID50病毒滴度是一个建立感染动物模型比较合适的剂量。

Objective To establish the Influenza H5N1 influenza virus infection ferrets animal model.Methods Groups of three 4-to 6-month-old ferrets were inoculated intranasally with 103 and 104TCID50 of H5N1 A /Vietnam/1203/2004 in a volume of 500uL.Ferrets were monitored daily for clinical signs of influenza infection,and body temperatures were recorded once daily.Every day take nasal turbinates for RT-PCR.On day 5 postinoculation,ferrets were euthanized and trachea,lung,heart,liver,slpeen,kidney,small intestine,and brain tissue were harvested,and examined by light microscopy.Results For each titer virus tested,in infected ferrets,we observed fever,weight loss,nasal rattling,sneezing and died.Virus was detected in the nasal turbinates and heart,liver,slpeen,kidney,small intestine,and brain from all of the iinfected ferrets.Of the ferrets inoculated with H5N1 virus,there show focal severe pneumonia in lung.Conclusions In clinical signs,virus detection,and histological changes were observed ferrets were infected H5N1 influenza virus,which 104 TCID50 titer was a more appropriate to establish flu animal models.