ICR小鼠感染呼肠孤病毒Ⅲ型的实验研究

摘要

Objective To establish reovirus type 3 (Reo-3) infection model of ICR mice and observe clinical syndrome,investigate virus load in target organs and serology dynamic change in infected mice.Method Animal adaption method was used to increase the virus's virulence.Systemic infection model was established with tail vein and intraperitoneally inoculation to ICR.The clinical sign was observed.The tissues and serums were harvested on 0 d before and 4 d、7 d、18 d、25 d、35 d、72 d、129 d after inoculation.Blood samples of three mice were randomly collected on 47 d、81 d、103 d.qPCR and ELISA were respectively used to detect the virus nucleic acid at target tissue and the antibody level.Results Virus virulence was increased after four times adaption trial.Mice death was observed at d7,d9,d10,d11,d16 post inoculation.The highest load of the virus was found in liver through qPCR,followed by heart,spleen,and lung.The peaks of viral load emerged from 6 to 11 d in all organs,negative results appeared at 129 d in the most organs.The earliest antibody detect time was on 18 d.The titer of antibody hit the peak on 25 d,then maintained a high level till to 129 d.Conclusion virus virulence can be enhanced significantly through mouse in-vivo inoculation,multiple organs inflammation and early death in mice can be identified by the systemic infection way of Reo-3.This experiment provides an approach to establish Reo-3 infected mice animal model and provides good experimental data for further research on Reo-3 infection mechanism.%目的 利用呼肠孤病毒Ⅲ型(Reo-3)感染ICR小鼠,分析小鼠的临床体征、靶器官病毒载量及血清抗体水平的动态变化.方法 通过动物体内适应试验增强病毒毒力,利用尾静脉注射与腹腔注射途径建立系统性感染ICR小鼠模型,观察动物临床体征,于感染0d、4d、7d、18d、25 d、35 d、72 d、129 d分别采集小鼠(2～3只小鼠)血液并剖检.47d、81d、103 d随机对3只小鼠进行血液采集跟踪.采用实时qPCR方法检测靶器官病毒核酸量,ELISA方法检测血清抗体水平.结果 病毒经过4次小鼠体内适应其毒力明显增强.小鼠攻毒后6d、7d、9d、10d、11d、16d出现死亡,实时qPCR结果显示,肝脏病毒载量最高,其次是心、脾、肺.多数器官病毒载量峰值出现在6～11d,129 d后,多数靶器官检测阴性.血清抗体最早在18d达到阳性,25 d达到峰值,并维持高抗体水平至试验结束.结论 小鼠体内适应方法可以明显增强Reo-3毒力,该病毒感染可引起小鼠多器官炎症反应,并导致小鼠死亡.本研究为该病毒模型建立及致病机理研究提供了参考数据.