Guo Yu-Fei, Cheng An-Chun, Wang Ming-Shu, Jia Ren-Yong, Wen Ming, Zhou Wei-Guang, Zhou Yi, Chen Xiao-Yue
Poultry Disease Research Center of Sichuan Agricultural University, Yaan 625014, China.
Bing Du Xue Bao. 2008 Sep;24(5):352-7.
The propagation characteristics of virulent duck plague virus (DPV) in duck embryo fibroblast (DEF) were studied by the method of light microscopy observation of DEF cell culture monolayer, electron microscopy observation of infected DEF cell culture, real-time PCR detecting virus propagation. The results demonstrated that on duck embryo fibroblast a number of plaques were formed by DPV 42 h postinfection. Electron microscopy of the ultrathin section of infected duck embryo fibroblasts demonstrated that the nucleic acid of DPV was round in shape with diameter of 35-45 nm and was often in a cluster in the nucleus of DEF. The nucleocapsid of DPV was round in shape with diameter of 90-100 nm and could be observed both in nucleus and cytoplasm of DEF. The mature DPV which had the structures of envelop and tegument was spherical in shape with diameter of 150-300 nm and was located in cytoplasmic vacuoles. DPV penetrated the DEF cell membrane by direct fusion between the viral envelop and the plasma membrane. Progeny viral nucleic acid was produced in the nucleus and the assembled nucleocapsids obtained the structure of tegument in the cytoplasm and obtained the structure of envelop by budding into the cytoplasmic vesicles. The mature DPV particles were released out of the cell through exocytosis of the cytoplasmic vesicles. Detection of DPV by real-time PCR demonstrated that virus in DEF began its obvious propagation 10 h postinfection and virus amount tended to increase until 30 h postinfection. DPV began to be released into the supernatant 22 h postinfection and the DPV amount peaked 50 h postinfection, when the virus content in DEF and supernatant both underwent approximately 10(3) fold increase. DPV mainly existed in the DEF and the virus content in DEF was 10(2)-10(3) fold than the supernatant.
通过鸭胚成纤维细胞（DEF）单层细胞培养的光学显微镜观察、感染DEF细胞培养的电子显微镜观察、实时荧光定量PCR检测病毒增殖的方法，研究了强毒鸭瘟病毒（DPV）在鸭胚成纤维细胞中的增殖特性。结果表明，感染DPV 42 h后，鸭胚成纤维细胞上形成了许多蚀斑。感染鸭胚成纤维细胞超薄切片的电子显微镜观察显示，DPV核酸呈圆形，直径为35 - 45 nm，常聚集在DEF细胞核内。DPV核衣壳呈圆形，直径为90 - 100 nm，在DEF的细胞核和细胞质中均能观察到。具有包膜和皮层结构的成熟DPV呈球形，直径为150 - 300 nm，位于细胞质空泡中。DPV通过病毒包膜与质膜的直接融合穿透DEF细胞膜。子代病毒核酸在细胞核中产生，组装好的核衣壳在细胞质中获得皮层结构，并通过出芽进入细胞质囊泡获得包膜结构。成熟的DPV颗粒通过细胞质囊泡的胞吐作用释放到细胞外。实时荧光定量PCR检测DPV表明，DEF中的病毒在感染后10 h开始明显增殖，病毒量在感染后30 h前呈上升趋势。DPV在感染后22 h开始释放到上清液中，感染后50 h DPV量达到峰值，此时DEF和上清液中的病毒含量均增加了约10³倍。DPV主要存在于DEF中，DEF中的病毒含量比上清液高10² - 10³倍。
