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当前位置: 首页 > 文章 > 新型鹅黄病毒JS804毒株的分离与鉴定 江苏农业学报 2011,27 (2) 354-360
Position: Home > Articles > Isolation and identification of a novel flavivirus strain JS804 in geese Jiangsu Journal of Agricultural Sciences 2011,27 (2) 354-360

新型鹅黄病毒JS804毒株的分离与鉴定

作  者：
黄欣梅;李银;赵冬敏;刘宇卓;张敬峰;何孔旺;侯继波;钮慧敏;李彬;温立斌;王永山;邵国青;张康宁;朱江宁;吴东明;高磊
单  位：
江苏省农业科学院兽医研究所;常州市四季禽业有限公司
关键词：
鹅;非序列依赖性单引物扩增法;黄病毒;JS804毒株
摘  要：
2010年4月至11月,江苏等地鸭、鹅发生了一种以产蛋率、采食量显著下降,出现脑炎样神经症状为特征的传染病。对发病鹅进行剖检观察,鸭胚尿囊腔途径接种发病鹅病料分离病原,电镜观察病毒粒子。对健康鹅接种分离病毒进行动物回归试验。在病毒核酸背景未知的情况下,应用非序列依赖性单引物扩增法结合DNA酶处理(DNase-sequence independent single primer amplification,DNase-SISPA)对病原基因进行扩增,并在此基础上设计了1对特异性引物对病毒基因进行PCR扩增。剖检结果发现病鹅的脑膜、肺脏、肝脏、心脏、卵巢等多器官出血,脾脏肿大坏死。含毒鸭胚尿囊膜超薄切片电镜观察显示:病毒粒子直径为50~60 nm。动物回归试验成功复制出该病并分离到病毒。应用DNase-SISPA方法发现了3个病毒相关基因片段,经序列比对分析,与黄病毒属(Flavi-virus)坦布苏病毒(Tembusu virus)基因序列有很高的同源性,分别为96%、88%、93%。据此设计特异性引物扩增出一段985 bp基因片段,该片段与黄病毒属坦布苏病毒E基因的核苷酸同源性为91%,氨基酸同源性为97%。将分离的鹅黄病毒毒株命名为Goose/Jiangsu/804/2010(简称JS804)。研究结果表明:此次鸭、鹅新发疫病的病原为一种新的黄病毒。
译  名：
Isolation and identification of a novel flavivirus strain JS804 in geese
作  者：
HUANG Xin-mei1,LI Yin1,ZHAO Dong-min1,LIU Yu-zhuo1,ZHANG Jing-feng1,HE Kong-wang1,HOU Ji-bo1,NIU Hui-min1,LI Bin1,WEN Li-bin1,WANG Yong-shan1,SHAO Guo-qing1,ZHANG Kang-ning2,ZHU Jiang-ning2,WU Dong-ming2,GAO Lei2(1.Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Key Laboratory of Animal Disease Diagnostic and Immunology,Ministry of Agriculture,National Center for Engineering Research of Veterinary Bio-products,Nanjing 210014,China;2.Changzhou Siji Poultry Co.,Ltd.,Jintan 213212,China)
关键词：
goose;sequence independent single primer amplification;Flavivirus;strain JS804
摘  要：
Since April 2010,a novel contagious disease in ducks and geese,with egg drop,feed uptake decline and nervous system signs,caused by a newly emerged virus has spread around Eastern China.Dissection conducted on the dead geese demonstrated haemorrhage in brain,lung,liver,heart,ovary,and enlarged and necrotic spleen.A virus strain isolated from geese in Jiangsu area was named Goose/Jiangsu/804/2010(JS804) strain.The virus was identified as an enveloped positive-stranded RNA virus with a size of approximately 50-60 nm in diameter.A similar disease was reproduced in healthy geese,and the virus was also successfully recovered from infected geese.After extraction of total DNA/RNA,DNase-SISPA method was used to amplify the genes of the virus.Three virus genome fragments were detected and sequenced,and the nucleotide sequences homologies with Tembusu virus,a member of the mosquito-borne virus cluster of the genus Flavivirus,Family Flaviviridae,were 96%,88%,and 93%,respectively.A 985 bp fragment was amplified with specific PCR primers,according to the gene sequence got with the DNase-SISPA method.The nucleotide sequence was 91%,and the amino acid sequence was 97% homologous to partial E gene of Tembusu virus.All the results indicated the causative agent of the new disease in ducks and geese occurring in China is a new flavivirus.

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