鸡IBV、NDV和AIV多重RT-PCR检测方法的初步建立

根据鸡传染性支气管炎病毒M基因、禽流感病毒NP基因和新城疫病毒F基因各设计1对引物,初步建立了针对鸡传染性支气管炎(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3对引物可对同一样品中的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333bp(IBV)、438bp(NDV)和196bp(AIV);建立的多重RT-PCR检测方法能够检测出1ng/μLAIV、1ng/μLNDV和10ng/μLIBV的cDNA,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3种鸡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 IBV, NDV and AIV

A multiple RT-PCR technology for diagnosis of IB,ND and AI has been constructed. And its specificity and sensitivity have also been tested. The results showed that IBV,NDV and AIV could be detected in one tube synchronously. The length of each specific product amplified by multiple RT-PCR was 333 bp (IBV),438 bp (NDV) and 196 bp (AIV),respectively. RNA contained 1 ng/μL AIV,1 ng/μL NDV and 10 ng/μL IBV can be tested by mRT-PCR,which demonstrated that the method had better specificity and sensitivity. In conclusion,the multi-PCR method can be used in diagnosis and epidemiological investigation of the above three virus respiratory diseases.