鸡ILV、IBV、NDV和AIV多重RT-PCR检测方法的初步建立

初步建立了鸡传染性喉气管炎(IL)、鸡传染性支气管炎(IB)、新城疫(ND)和禽流感(AI)的多重RT- PCR鉴别诊断方法,并对其特异性和敏感性进行了检测。结果表明,所设计的4对引物可对同一样品中的ILV、IBV、NDV和AIV进行特异性扩增,所扩增的目的片断的长度分别为620 bp(ILV)、445 bp(IBV)、344 bp(NDV)和240 bp (AIV);建立的RT-PCR检测方法能够检测出10 pg AIV、1 ng NDV和10 ng IBV的RNA及100 pg ILV的DNA,说明该检测方法特异性强,敏感性较高。本研究所建立的多重RT-PCR方法可用于上述鸡4种病毒性呼吸道疾病的快速鉴别诊断,在临床诊断和流行病学调查方面具有较好的应用前景。

Initial establishment of multiple RT-PCR for detecting ILV、IBV、NDV and AIV

A multi-PCR technology for IL,IB,ND and AI identity ans diagnosis has been constructed. And its specificity and sensitivity have also been tested . The results show that ILV, IBV, NDV and AIV could be detected in one tube synchronously. The length of each specific product amplified by multi-PCR is 620 bp (ILV ) ,445 bp (IBV),344 bp (NDV),and 240 bp (AIV),respectively. RNA contained in 10 pg AIV,1 ng NDV and 10 ng IBV and DNA contained in 100 pg ILV also can be tested by RT-PCR,which demonstrates that the method has better specificity and sensitivity. In a word, we have constructed an efficient multi-PCR method for diagnosing and contyocing the above four virus respiratory diseases.