Taura syndrome virus(TSV) is one of the important pathogenic agents of Penaeus vannami,which causes prawn infection with serious disease in China in recent years.245 samples collected from 26 farms of Zhangzhou,Xiamen...Taura syndrome virus(TSV) is one of the important pathogenic agents of Penaeus vannami,which causes prawn infection with serious disease in China in recent years.245 samples collected from 26 farms of Zhangzhou,Xiamen,Shenzhen,Yangjiang,Ningbo and Guangzhou.were detected for TSV with RT-PCR,the positive rates were 100%,33.3%,40.7%,50% and 40%,respectively.Southern blotting and nucleotide sequencing were performed to confirm the specificity of the amplified RT-PCR products.The homology of 231bp nucleotide sequences reached 98.3%-100% when analysed and compared with the GenBank using the BLAST program.Phylogenetic analyses clustered the TSV isolates into two groups: one contained USA and Chinese Taiwan isolates;the other contained Vietnam isolate(YN1) and 21 isolates of China's Mainland.The 21 China isolates clustered into one branch,while the YN1 distributed in another branch.5 strains from Guangzhou and 2 strains from Shenzhen constituted one subcluster of the China branch.展开更多
根据对虾桃拉综合征病毒(TSV)结构蛋白基因保守序列,利用Primer Explorer 4.0软件设计了4条引物,建立桃拉综合征病毒环介导逆转录等温扩增快速(RT-LAMP)检测方法,对反应温度和反应时间等参数进行了优化;同时将建立的RT-LAMP检测方法与RT...根据对虾桃拉综合征病毒(TSV)结构蛋白基因保守序列,利用Primer Explorer 4.0软件设计了4条引物,建立桃拉综合征病毒环介导逆转录等温扩增快速(RT-LAMP)检测方法,对反应温度和反应时间等参数进行了优化;同时将建立的RT-LAMP检测方法与RT-PCR进行比较分析,RT-LAMP的检测灵敏度比RT-PCR高10倍。结果表明,RT-LAMP最适反应在60℃恒温条件60 min内完成,凝胶电泳呈现梯型条带;反应体系中添加SYBR Green I荧光染料后,绿色的阳性结果明显区别于橙色阴性结果;RT-LAMP对感染TSV的对虾检测呈阳性,而对对虾其他常见病原检测均为阴性,具有高的特异性。展开更多
文摘Taura syndrome virus(TSV) is one of the important pathogenic agents of Penaeus vannami,which causes prawn infection with serious disease in China in recent years.245 samples collected from 26 farms of Zhangzhou,Xiamen,Shenzhen,Yangjiang,Ningbo and Guangzhou.were detected for TSV with RT-PCR,the positive rates were 100%,33.3%,40.7%,50% and 40%,respectively.Southern blotting and nucleotide sequencing were performed to confirm the specificity of the amplified RT-PCR products.The homology of 231bp nucleotide sequences reached 98.3%-100% when analysed and compared with the GenBank using the BLAST program.Phylogenetic analyses clustered the TSV isolates into two groups: one contained USA and Chinese Taiwan isolates;the other contained Vietnam isolate(YN1) and 21 isolates of China's Mainland.The 21 China isolates clustered into one branch,while the YN1 distributed in another branch.5 strains from Guangzhou and 2 strains from Shenzhen constituted one subcluster of the China branch.
文摘根据对虾桃拉综合征病毒(TSV)结构蛋白基因保守序列,利用Primer Explorer 4.0软件设计了4条引物,建立桃拉综合征病毒环介导逆转录等温扩增快速(RT-LAMP)检测方法,对反应温度和反应时间等参数进行了优化;同时将建立的RT-LAMP检测方法与RT-PCR进行比较分析,RT-LAMP的检测灵敏度比RT-PCR高10倍。结果表明,RT-LAMP最适反应在60℃恒温条件60 min内完成,凝胶电泳呈现梯型条带;反应体系中添加SYBR Green I荧光染料后,绿色的阳性结果明显区别于橙色阴性结果;RT-LAMP对感染TSV的对虾检测呈阳性,而对对虾其他常见病原检测均为阴性,具有高的特异性。
