气管插管法接种H5N1病毒感染恒河猴及系统组织病毒检测

摘要

目的 测试气管插管法接种高致病性禽流感病毒H5N1感染恒河猴的优势效果及疾病分析,为有效感染恒河猴、制备H5N1疾病模型提供实验依据.方法 使用人源H5N1病毒液经气管插管滴入恒河猴上呼吸道进行感染,观察感染恒河猴的临床表现,每天采集咽拭子、鼻灌洗液,在感染前2d感染后第3、5、7天采血,感染后第3和7天分别解剖1只恒河猴,取支气管淋巴结、肠淋巴结、鼻甲、心、肝、脾、肺、肾、肠、气管、脑及血液进行病毒分离、核酸载量检测和血常规测定.结果 感染后第2天恒河猴出现食欲下降,活动减少,并伴有一过性体温升高,白细胞数和淋巴细胞数下降.咽拭子、鼻灌洗液、肺、心、气管、脑、肝、肾、肠和血液中都能分离到H5N1病毒.结论 气管插管法接种H5N1病毒能有效感染恒河猴,并在猴体内多组织中分离、检测到病毒,为制备完善的H5N1模型和检测指标确定、进一步研究H5N1病毒的致病机制等奠定了基础.%Objective To evaluate the feasibility of endotracheal intubation route in preparation of H5N1 virus infection in rhesus macaques and to obtain a more sensitive animal model. Methods The rhesus macaques were infected with H5N1 avian influenza virus by endotracheal intubation. Clinical signs were assessed daily, and throat swab, nasal wash , major organs and blood were collected for detection of virus load, isolation and blood routine analysis. Results After infection the animals exhibited anorexia, lethargy and transient fever, and the total number of circulating leukocytes and lymphocytes was decreased. Virus isolation, RT-PCR assay of the throat swabs, nasal washs materials showed that the intestine, lung, brain, spleen, kidney, heart and liver were the target of the virus replication and the virus could replicate in organs outside the lung. In addition, virus was also detected in the blood 3 days after infection. Conclusions Rhesus macaques can be infected with H5N1 virus by endotracheal intubation, and it is proved to be a good method to obtain a more sensitive animal model of H5N1 virus infection. The experimental results will lay the foundation for further studies of pathogenetic mechanism of H5N1 infection.