重组酶聚合酶扩增技术结合胶体金试纸条检测新城疫病毒方法的建立及应用  

Establishment and application of recombinant enzyme polymerase chain reaction combined with colloidal gold strip for detection of Newcastle disease virus

作　　者：唐豪[1] 赵子惠[3] 张登基 贺泂杰[2] TANG Hao;ZHAO Zi-hui;ZHANG Deng-ji;HE Jiong-jie(The Animal Pestilence Prevention and Control Center of Gansu Province,Lanzhou 730000,China;Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS,Lanzhou 73005,China;Gansu Institute of Animal and Veterinary Sciences,Pingliang744000,China)

机构地区：[1]甘肃省动物疫病预防控制中心,甘肃兰州7300003 [2]中国农业科学院兰州畜牧与兽药研究所,甘肃兰州73005 [3]甘肃省畜牧兽医研究所,甘肃平凉744000

出　　处：《中国兽医科学》2023年第3期298-303,共6页Chinese Veterinary Science

基　　金：甘肃省科技计划项目-重点研发计划(21YF5NA143);甘肃省农业生物技术专项(2015-84)。

摘　　要：本研究旨在根据鸡新城疫病毒(Newcastle disease virus)F基因序列,利用Oligo 7设计RPA引物以及nfo探针,建立快速检测新城疫病毒的结合胶体金试纸条的RPA检测方法。在试验中,对探针浓度、引物浓度、反应温度、反应时间进行了优化;并将检测结果与荧光RT-PCR商品化试剂盒的检测结果进行了比较。结果显示,本方法采用50μL体系,37℃20 min即可对新城疫病毒F基因进行快速检测。该方法灵敏度高,特异性强,重复性好,检测结果与荧光RT-PCR商品化检测试剂盒检测结果一致,为养殖场检测新城疫病毒提供了一种简便可靠的检测方法。In this study,according to the F gene of Newcastle disease virus(NDV),we designed RPA primers and nfo probes using oligo 7,to establish a RPA combined with colloidal gold strip for the detection of NDv.The concentrations of probes and primer,reaction temperature and reaction time were optimized.The detection result using the RPA was compared with the detection result using the fluorescent RT-PCR commercial detection kit.In result,this method used 50μL system,and the NDV F gene could be rapidly detected at 37 C for 20 minutes.This method has high sensitivity,strong specificity,good repeatability,and the detection results are consistent with those of the fluorescent RT-PCR commercial detection kit.In conclusion,it provides a simple and reliable method for the detection of NDV on aquaculture farms.

关 键 词：重组酶聚合酶扩增技术(RPA) 鸡 新城疫病毒 胶体金试纸条 

分 类 号：S852.659.5[农业科学—基础兽医学]