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树鼩是灵长类动物的近亲,且具有体型小、繁殖周期短、饲养管理成本低等优点,长期以来被认为有望替代灵长类动物用于人类疾病的动物模型研究。该文研究了肠道病毒EV71对幼年中缅树鼩的感染特点,探索建立EV71感染树鼩动物模型及替代灵长类动物的可行性。实验分别采用灌胃、滴鼻和尾静脉注射3种方式,感染3月龄树鼩,定期观察动物临床症状和血常规变化和定期采集血液和粪便样品,并使用中和抗体试验、reverse transcription-PCR(RT-PCR)和Real-TimePCR等技术,检测相关样品中和抗体效价、病毒核酸及载量变化。结合组织病理学检查,分析感染病变特点。研究结果发现,实验组树鼩体温在第4天前后开始升高,白细胞、淋巴细胞也有类似趋势;3种攻毒方式均可检测到病毒载量,峰值出现在第10天前后,灌胃途径尤为明显,血清的最大中和抗体效价为1:16。感染EV71病毒的树鼩2周左右出现急性松弛性瘫痪,大体解剖发现伴有尿潴留症状,组织病理学检查发现在脑、肺、肠、脾脏、肾脏等部位发生病变。结果表明,EV71病毒通过灌胃、滴鼻和静脉注射3种方式均可以感染中缅树鼩,并使之出现神经系统及相关病变,该实验为将来研究EV71感染树鼩致病机理、建立手足口病树鼩动物模型奠定了理论基础。
Tree shrews are close relatives of primates with the advantages of small size, short reproductive cycle, and low cost of feeding and management. They have long been considered to be promising animal models for substituting primate for human diseases. In this paper, the infection characteristics of intestinal virus EV71 in juvenile tree shrews were studied, and the feasibility of establishing animal model of shrews of EV71 infection and replacing primates was explored. The experiment was carried out by gavage, intranasal and tail vein injection of 3 ways, 3-month-old tree shrew infection, regular observation of clinical symptoms and blood changes in animals and blood and stool samples regularly collected and neutralizing antibody test, reverse transcription- PCR (RT-PCR) and Real-TimePCR and other technologies to detect the neutralizing antibody titer, viral nucleic acid and load changes. Combined with histopathological examination, analysis of the characteristics of infected lesions. The results showed that in the experimental group, the body temperature of tree shrews began to increase around day 4, and the leukocytes and lymphocytes also showed similar trends. The viral load was detected in all three kinds of challenge modes, with the peak value occurring around the 10th day and the gavage route Most notably, the serum maximum neutralizing antibody titer was 1:16. In the tree shrew infected with EV71 virus, acute flaccid paralysis occurred for about 2 weeks. The gross anatomy was found to be associated with the symptoms of urinary retention. Histopathological examination revealed lesions in the brain, lung, intestine, spleen and kidney. The results showed that the EV71 virus could infect CNS and its related pathological changes through gavage, intranasal instillation and intravenous injection. In this experiment, the pathogenic mechanism of EV71 infection in tree shrew was studied in the future, Stump disease tree animal model laid the theoretical foundation.