基因Ⅵ型新城疫病毒荧光RT-PCR检测方法的建立与应用  

Establishment and Application of the Fluorescent RT-PCR for Detecting Newcastle Disease Virus Genotype Ⅵ

作　　者：王静静[1] 于晓慧 克军宏 彭真奇 邢安琪 刘华雷[1] Wang Jingjing;Yu Xiaohui;Ke Junhong;Peng Zhenqi;Xing Anqi;Liu Hualei(China Animal Health and Epidemiology Center,Qingdao,Shandong 266032,China;Tarim University,Alaer,Xinjiang 843300,China;Anhui Agricultural University,Hefei,Anhui 230036,China;Ningxia University,Yinchuan,Ningxia 750021,China)

机构地区：[1]中国动物卫生与流行病学中心,山东青岛266032 [2]塔里木大学,新疆阿拉尔843300 [3]安徽农业大学,安徽合肥230036 [4]宁夏大学,宁夏银川750021

出　　处：《中国动物检疫》2022年第6期114-118,共5页China Animal Health Inspection

基　　金：国家重点研发计划项目(2016YFD0500800)。

摘　　要：鸽新城疫是严重危害鸽业健康发展的主要疫病之一,近年来流行呈上升趋势。为满足开展基因Ⅵ型鸽源新城疫病毒快速检测的迫切需求,针对国内流行的基因Ⅵ型新城疫病毒F基因保守区域设计特异性引物和探针,建立了一种可特异检测基因Ⅵ型新城疫病毒的实时荧光RT-PCR方法,并对该方法的特异性、敏感性进行了评估。结果显示,该方法与其他基因型新城疫病毒和常见禽病病毒无交叉反应,检测下限为5 copies/μL。利用该方法对临床采集的120份鸽口咽/泄殖腔拭子样品进行检测,检测结果与病毒分离结果完全一致。结果表明,本研究建立的实时荧光RT-PCR方法特异性强、敏感性高、准确性好,可用于基因Ⅵ型新城疫病毒的快速检测。Pigeon Newcastle disease is one of the major diseases seriously endangering the healthy development of pigeon industry,which has been increasingly prevalent in recent years. In order to meet the urgent needs for rapid detection of Newcastle disease virus genotype Ⅵ(NDV-Ⅵ)in pigeons,the specific primers and probes were designed based on the conserved region of F gene of NDV-Ⅵ,and a fluorescent RT-PCR for specifically detecting the virus was established,followed by evaluation on its specificity and sensitivity. It was showed that no cross-reaction with other NDVs and common poultry viruses was observed,and the detection limit was 5 copies/μL. 120 throat and cloacal swabs collected from pigeons were detected by this method,and the results were consistent with those of virus isolation. It was concluded that the established method was highly specific,sensitive and accurate,which could be used to rapidly detect NDV-Ⅵ.

关 键 词：基因Ⅵ型新城疫病毒 实时荧光RT-PCR 检测 

分 类 号：S855.3[农业科学—临床兽医学]