Establishment of a novel and highly permissive cell line for the efficient replication of cyprinid herpesvirus 2 (CyHV-2).
作者： Jie Ma , Nan Jiang , Scott E. LaPatra , Ling Jin , Jin Xu
DOI: 10.1016/J.VETMIC.2015.04.006
关键词: Immunofluorescence 、 Virology 、 Cytopathic effect 、 Cell 、 Fluorescence in situ hybridization 、 Cell fusion 、 Carp 、 Biology 、 Virus 、 Cell culture
摘要: Abstract Haematopoietic necrosis of gibel carp (Carassius auratus gibelio) is caused by cyprinid herpesvirus 2 (CyHV-2) and has huge economic losses in aquaculture worldwide. Currently the isolation propagation CyHV-2 vitro very difficult due to lack permissive cell lines. Studies on pathogenesis have been hampered because virus not extensively characterized. In this study, a novel line from brain carp, denoted GiCB, established Sustainable GiCB confirmed infection titration, PCR, transmission electron microscopy, immunofluorescence assay fluorescence situ hybridization. The cells showed typical cytopathic effect day 6 post-infection with including shrinkage, rounding, fusion cytoplasmic vacuolization. titer reached 107.5±0.37TCID50/ml successfully passaged over 50 times line. Electron microscopy analysis revealed complete replication cells. CyHV-2-infected reacted strongly polyclonal antibodies against RNA hybridized specifically probes. Additionally, an experimental demonstrated that produced 100% mortality carp. All results provide solid evidence highly for CyHV-2. This significant advancement will promote additional research fish future.
elsevier.com
sci-hub.se 参考文章(35)
蒋一珪, 梁绍昌, 桂建芳, 朱蓝菲, Discovery of Multiple Tetraploids in Artificially Propagated Populations of Allogynogenetic Silver Crucian Carp and Their Breeding Potentialities Chinese Science Bulletin. ,vol. 38, pp. 327- 331 ,(1993) , 10.1360/SB1993-38-4-327
U. Kunz, M. Neukirch, Isolation and preliminary characterization of several viruses from koi (Cyprinus carpio) suffering gill necrosis and mortality. Bulletin of The European Association of Fish Pathologists. ,vol. 21, pp. 125- 135 ,(2001)
J. Ma, S. Sun, L. Zeng, Y. Lu, Establishment, characterization and viral susceptibility of two cell lines derived from leopard wrasse Macropharyngodon geoffroy. Journal of Fish Biology. ,vol. 83, pp. 560- 573 ,(2013) , 10.1111/JFB.12193
Lesley Graham, Jan Marc Orenstein, Processing tissue and cells for transmission electron microscopy in diagnostic pathology and research Nature Protocols. ,vol. 2, pp. 2439- 2450 ,(2007) , 10.1038/NPROT.2007.304
N. C. Bols, L. E. J. Lee, Technology and uses of cell cultures from the tissues and organs of bony fish. Cytotechnology. ,vol. 6, pp. 163- 187 ,(1991) , 10.1007/BF00624756
S Miwa, T Ito, M Sano, Morphogenesis of koi herpesvirus observed by electron microscopy. Journal of Fish Diseases. ,vol. 30, pp. 715- 722 ,(2007) , 10.1111/J.1365-2761.2007.00850.X
J. Braunwald, H. Nonnenmacher, F. Tripier-Darcy, Ultrastructural and biochemical study of frog virus 3 uptake by BHK-21 cells. Journal of General Virology. ,vol. 66, pp. 283- 293 ,(1985) , 10.1099/0022-1317-66-2-283
C.M. Wen, C.W. Lee, C.S. Wang, Y.H. Cheng, H.Y. Huang, Development of two cell lines from Epinephelus coioides brain tissue for characterization of betanodavirus and megalocytivirus infectivity and propagation Aquaculture. ,vol. 278, pp. 14- 21 ,(2008) , 10.1016/J.AQUACULTURE.2008.03.020
Jie Ma, Lingbing Zeng, Yuding Fan, Yong Zhou, Nan Jiang, Qian Chen, Significant inhibition of two different genotypes of grass carp reovirus in vitro using multiple shRNAs expression vectors Virus Research. ,vol. 189, pp. 47- 55 ,(2014) , 10.1016/J.VIRUSRES.2014.05.009
Xiaohong Huang, Youhua Huang, Zhengliang Ouyang, Lixiao Xu, Yang Yan, Huachun Cui, Xin Han, Qiwei Qin, Singapore grouper iridovirus, a large DNA virus, induces nonapoptotic cell death by a cell type dependent fashion and evokes ERK signaling Apoptosis. ,vol. 16, pp. 831- 845 ,(2011) , 10.1007/S10495-011-0616-Y