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鸡ILV、IBV、NDV和AIV多重RT-PCR检测方法的初步建立
王 非1,2, 蒋建一3, 华炯刚2
1.安徽农业大学 动物科技学院;2.浙江农业科学院 病毒学与生物技术研究所;3.杭州市畜牧兽医总站
摘要:
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。
关键词:  多重PCR  鸡传染性喉气管炎  鸡传染性支气管炎  鸡新城疫  禽流感  鉴别诊断
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基金项目:杭州市科技局项目(20051332B26)
Initial establishment of multiple RT-PCR for detecting ILV、IBV、NDV and AIV
Abstract:
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.
Key words:  multiplex PCR,infectious laryngotracheitis virus,infectious bronchitis virus,newcastle diseases virus,avin infectious virus,identity and diagnosis

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