鸡新城疫病毒RT-PCR检测方法的建立及应用  

Establishment and Application of RT-PCR Technigue for Detection of Newcastle Disease Virus

作　　者：陈贤德 汪洋 

机构地区：[1]青海省海西州茫崖行委动物卫生监督所,青海海西816400 [2]山东滨州畜牧兽医研究院,山东滨州256600

出　　处：《动物医学进展》2013年第6期208-211,共4页Progress In Veterinary Medicine

摘　　要：根据GenBank上发表的鸡新城疫病毒(NDV)基因组中F基因的保守序列,利用Primer 5.0设计一对引物,扩增201bp特异性核酸片段,建立了检测NDV的RT-PCR方法。特异性试验结果表明,针对NDV能扩增出201bp的核酸片段,而对传染性支气管炎病毒(IBV)、禽流感病毒(AIV)均无特异性条带出现。敏感性试验结果表明,该方法的最低检出量为10pg的模板。利用该方法对10株从青海、内蒙、吉林及河北分离的疑似鸡NDV毒株进行检测,结果均为阳性。上述结果表明,本试验建立的RT-PCR方法特异性强、稳定性好。该方法的建立为NDV的检测及流行病学调查提供了可靠的手段。According to the sequences of Newcastle disease virus F gene published in GenBank,a pair of primers were designed and synthesized to specifically amplify the 201 bp nucleotide fragment,the RT-PCR technique for detection of NDV was established.The 201 bp special strip was found only in the PCR amplification of the NDV in the specificity assay,there were no the same strip appeared in the other viruses.The sensitivity test result indicated that as little as 10 Pg in sample can be detected in PCR.All of the ten uncertain isolates detected by the established RT-PCR technique were positive.The above results showed that the technique provided a more sensitive,specific and rapid method for diagnosis and epidemiological survey of NDV.

关 键 词：新城疫病毒 F基因 逆转录-聚合酶链反应 

分 类 号：S852.659.5[农业科学—基础兽医学]