二重PCR检测鸭瘟病毒和鹅细小病毒的初步研究

Developmemt of duplex PCR assay for detection of duck plague virus and goose parvovirus

根据鸭瘟病毒(Duck plague virus,DPV)DNA聚合酶基因和鹅细小病毒(Goose parvovirus,GPV)NS基因序列分别设计2对引物,应用这2对引物对混合样品中鸭瘟病毒和鹅细小病毒进行了二重PCR扩增。PCR产物的琼脂糖凝胶电泳结果表明2条特异性带为563bp(DPV)和1146bp(GPV),与实验设计相符,且与常规单一PCR结果一致。二重PCR反应检测出的DPV含量相当于20个PFU;检测出的GPV含量相当于0.1个ELD50,对禽痘病毒和减蛋综合征病毒的核酸未能扩增出任何条带,特异性良好。该二重PCR能够在一次扩增反应中检测两种病毒,为两种鹅病的病原检测和诊断提供了一种简便、经济、快速的手段。

A duplex PCR assay was developed using 2 pairs of primers designed based on the conserved region of duck plague virus （DPV） DNA polyrnerase gene and goose parvovirus NS gene, Two specific DNA products 563 bp of DPV and 1 146 bp of GPV were simultaneously amplified from the viruses. The assay could detect 20 PFU of DPV and 0.1 ELD50 of GPV and did not amplify any PCR products from FPV and EDSV controls. The result suggested that the duplex PCR is a specific and sensitive method which could be used for detection and diagnosis of latent and clinical DPV and GPV infections,