鸭坦布苏病毒RT-PCR检测方法的建立及应用

Establishment and Application of RT-PCR Detective Method for Duck Tembusu Virus

关键词:  鸭坦布苏病毒  E基因  RT-PCR

Key words:  Duck Tembusu virus  E gene  RT-PCR

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。

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.