鸭新城疫病毒与番鸭细小病毒二重PCR方法的建立

根据GenBank中新城疫病毒(NDV)L基因和番鸭细小病毒(MDPV)Vp3基因的保守序列,采用Primer Premier 5.0软件设计并合成了2对引物。通过优化反应条件及特异性、敏感性评价,建立了能同时检测鸭源NDV和MDPV的二重PCR方法。该方法对鸭源NDV和MDPV的检测敏感性分别为30和16 fg。同时使用该方法对鸭瘟病毒、鸭肝炎病毒、鸭圆环病毒、H9亚型禽流感病毒、鸭黄病毒、沙门氏菌、大肠杆菌和禽多杀性巴氏杆菌进行检测,结果显示全为阴性。本研究建立的鸭源NDV和MDPV的二重PCR检测方法,具有特异、敏感、快速、重复性好等优点,可用于鸭源NDV和MDPV感染的快速鉴别检测。

Establishment of a Duplex PCR Assay for Detection of Duck Newcastle Disease Virus and Muscovy Duck Parvovirus

According to the gene sequences of duck Newcastle disease virus (NDV) L gene and Muscovy duck parvovirus (MDPV) Vp3 gene in GenBank, two sets of specific primers were designed by Primer Premier 5.0. The duplex PCR assay was developed through optimization of reaction conditions and validation of specificity, sensitivity and repetitiveness of the method. The sensitivity of the assay was 30 fg for NDV and 16 fg for MDPV. No specific bands of the same sizes were amplified from other duck pathogens, such as duck plague virus, duck hepatitis virus, duck circovirus, H9 subtype avian influenza virus, duck flavivirus, Salmonella, E.coli, avian Pasteurella multocida. This duplex PCR assay was a quick, sensitive and specific test for detection of duck NDV and MDPV, and would be useful for the control of these viruses in ducks.