番鸭呼肠孤病毒套式RT-PCR检测方法的建立及应用

根据GenBank上发表的鸭呼肠孤病毒基因组序列,利用生物学软件设计合成内外2对引物,建立了检测番鸭呼肠孤病毒(Muscovy duck reovirus,MDRV)的套式RT-PCR检测方法,并运用建立的检测方法对分离病毒与其他禽病病毒进行检测。结果显示,该方法能从MDRV中扩增到与预期大小相符的特异性目的片段,检测灵敏度达到0.1 pg病毒RNA,对禽呼肠孤病毒(avian reovirus,ARV)、鸡传染性法氏囊病病毒(infectious bursal disease virus,IBDV)、番鸭细小病毒(Muscovy duck parvovirus,MDPV)、鹅细小病毒(goose parvovirus,GPV)、鸭病毒性肝炎病毒(duck hepatitis virus,DHV)等病毒样品的扩增结果均为阴性。因此,本研究为番鸭呼肠孤病毒病的快速检测及诊断研究提供参考。

Establishment and Application of A Nest RT-PCR Method for Detection of Muscovy Duck Reovirus

According to the GenBank login duck reovirus genome sequence, using biology software to design and synthesis two pairs of primers, established a nest RT-PCR detection method for the detection of Muscovy duck reovirus(MDRV), and used the detection method of the isolated virus and other poultry virus for application testing. The results showed that the method could amplify specific target fragment from MDRV, sensitivity to 0.1 pg virus RNA, while other viruses of avian reovirus (ARV), chicken infectious bursal disease virus (IBDV), Muscovy duck parvovirus(MDPV), goose parvovirus (GPV), duck hepatitis virus (DHV) and sample amplification results were negative. Therefore, the research provided a reference for MDRV disease detection and diagnose.