YN87448 virus strain was isolated from a fevered female patient (52 years old ) in Yunnan Province in 1986, and was identified as a member of Alphavirus using serological method. One primer was designed from common ha...YN87448 virus strain was isolated from a fevered female patient (52 years old ) in Yunnan Province in 1986, and was identified as a member of Alphavirus using serological method. One primer was designed from common hairpin conserved region of Alphavirus. Two fragments were amplified by single primer differential RT PCR, and they were cloned into pGEM T vector. Sequences were determined, and then analyzed by Software and GenBank. Results showed that the 1118pb long fragment was highly homologous to the sequence of Sindbis like virus S.A.A.R86. The homogeneity of the 1118bp fragment between YN87448 virus strain and S.A.A.R86 virus strain was 98%. Single primer differential RT PCR is a useful method with simple, economic, and practical value for Alphavirus identification.展开更多
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...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。展开更多
文摘YN87448 virus strain was isolated from a fevered female patient (52 years old ) in Yunnan Province in 1986, and was identified as a member of Alphavirus using serological method. One primer was designed from common hairpin conserved region of Alphavirus. Two fragments were amplified by single primer differential RT PCR, and they were cloned into pGEM T vector. Sequences were determined, and then analyzed by Software and GenBank. Results showed that the 1118pb long fragment was highly homologous to the sequence of Sindbis like virus S.A.A.R86. The homogeneity of the 1118bp fragment between YN87448 virus strain and S.A.A.R86 virus strain was 98%. Single primer differential RT PCR is a useful method with simple, economic, and practical value for Alphavirus identification.
文摘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。
