犬副流感病毒实时荧光RPA检测方法的建立  

Establishment of real-time recombinase polymerase amplification assay for the detection of canine parainfluenza virus

作　　者：李春华[1] 魏晓锋 熊炜 薛俊欣 黄忠荣 李健 LI Chun-hua;WEI Xiao-feng;XIONG Wei;XUE Jun-xin;HUANG Zhong-rong;LI Jian(Institute of Animal Husbandry and Veterinary Science , Shanghai Academy of Agricultural Sciences , Shanghai 201106, China;Shanghai Laboratory Animal Research Center,Shanghai 201203, China;Shanghai Customs District, Shanghai 200135, China)

机构地区：[1]上海市农业科学院畜牧兽医研究所,上海201106 [2]上海实验动物研究中心,上海201203 [3]上海海关,上海200135

出　　处：《中国兽医科学》2019年第10期1233-1238,共6页Chinese Veterinary Science

基　　金：国家质量基础的共性技术研究与应用专项(2017YFF0210202);上海市科委实验动物研究专项(17140900300)

摘　　要：为快速监控犬猫科动物中犬副流感病毒(CPIV)的感染情况,本研究针对CPIV核衣壳蛋白基因的保守序列,设计特异性引物和荧光探针,建立了CPIV实时荧光重组酶聚合酶等温扩增方法(real-time RPA)。结果表明,该方法具有良好的特异性,与其他犬病毒和同属的副黏病毒均未出现交叉反应;其敏感性与RT-PCR方法相近;将该方法应用于CPIV感染犬的临床组织样本、体表样本的检测,证实该实时荧光RPA方法具有良好的稳定性和可靠性。与现有的核酸检测方法相比,该实时荧光RPA方法检测周期仅30min即可完成结果判读,满足了犬猫科动物CPIV快速检测的需要。In order to accelerate inspection efficiency of CPIV,a real-time recombinase polymerase amplification(real-time RPA) was developed by using pairs of specific primers and a probe against conserved sequence of nucleocapsid gene of CPIV.The results showed that the new method had good specificity to CPIV and there was no cross-reaction to other canine virus and other paramyxovirus.The real-time RPA method had the same sensitivity as RT-PCR method,and its excellent stability was confirmed by detection of CPIV infected tissue and clinical samples.Compared with other detection methods of nucleic acid,the real-time RPA could finish detection within 30 min.It meets the requirement of rapid detection of CPIV among canid and felid.

关 键 词：犬 犬副流感病毒 实时荧光RPA方法 

分 类 号：S852.659.5[农业科学—基础兽医学]