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鸭坦布苏病毒RT-PCR检测方法的建立及应用
邹敏,张青,王红,刘东,刘新文,范根成,吴发兴
0
(青岛易邦生物工程有限公司)
摘要:
为建立一种能快速检测鸭坦布苏病毒(Tembusu virus,TMUV)的分子生物学方法,从GenBank中下载了51个具有代表性的黄病毒全基因组序列及白洋淀病毒等5个禽源黄病毒株E基因序列,进行了黄病毒E基因序列分析,应用Primer 5.0软件设计了一对引物,扩增目的片段为549 bp,进行了TMUV RT-PCR检测方法的特异性、敏感性、重复性试验,建立了TMUV RT-PCR检测方法应用该方法对收集的14份临床疑似蛋鸭鸭坦布苏病毒病发病样品进行了检测,共检出6份TMUV核酸阳性样品,检出率达42.86%。结果表明,该方法可用于水禽坦布苏病毒病的临床快速诊断、检测及流行病学调查。
关键词:  鸭坦布苏病毒  E基因  RT-PCR
DOI:
投稿时间:2018-09-05修订日期:2018-11-01
基金项目:青岛市动物保健品行业智库联合基金(16-10-1-1-jch)
Establishment and Application of RT-PCR Detective Method for Duck Tembusu Virus
(YEBIO Bioengineering Co.,Ltd of Qing dao)
Abstract:
To provide a molecular biological method for rapid detection of the duck Tembusu virus(TMUV), the complete genome sequence of 51 representative Flavivirus ,E gene of the BYD virus and 5 Avian Flavivirus were downloaded from the GenBank,multiple sequences analysis of Flavivirus E gene was carried out, and using the Primer 5.0 software,a pair of primers were designed,the motive fragment of amplification was 549 bp. A Tembusu virus RT-PCR detective method was established. The method had some advantages such as fast,specific,sensitive and repeatable.The method was used to detect 14 suspected clinic samples. There were 6 TMUV positive samples, the detection rate was 42.86%. The results showed that this method could be used for the rapid diagnosis, detect and epidemiological investigation of the waterfowl Tembusu virus disease.
Key words:  Duck Tembusu virus  E gene  RT-PCR

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