鸭出血性卵巢炎病毒RT-PCR检测方法的建立

Establishment of RT-PCR for detecting duck hemorrhagic ovaritis causing abrupt egg-laying reduction in ducks

摘要: 参照GenBank上登录的黄病毒科黄病毒属NS5基因片段序列设计引物,建立能检测引起鸭出血性卵巢炎病毒的RT-PCR方法。该方法能从鸭出血性卵巢炎病毒扩增出1条约470 bp的特异性片段,从H9亚型禽流感病毒、鸭呼肠孤病毒、鸭瘟病毒、鸭肝炎病毒、鸭源禽Ⅰ型副粘病毒均不能扩增出目的片段;敏感性试验显示建立的RT-PCR方法最低可检出20 pg病毒核酸。以上结果表明建立的RT-PCR方法具有较好的特异性、敏感性和准确性,可用于该病的临床诊断和流行病学调查。

Abstract: A pair of primers based on NS5 gene sequences of different strains of genus Flavivirus was designed for amplification of 470 bp fragments.No positive target fragment was available from H9 subtype influenza virus,Muscovy duck reovirus,duck plague,duck hepatitis virus(DHV-Ⅰ) or duck-origin avian paramyxovirus typeⅠ.The detection limit reached 20 pg under the optimized conditions.The results showed that the RT-PCR was sensitive,specific and applicable for rapid laboratory diagnosis and epidemiologic surveillance.