微滴数字RT-PCR检测南芥菜花叶病毒  

Establishment of droplet digital RT-PCR assay for detection of arabis mosaic virus

作　　者：赵晓丽 杨术江 陈红运 向军 张永江[4] 邓丛良 Zhao Xiaoli;Yang Shujiang;Chen Hongyun;Xiang Jun;Zhang Yongjiang;Deng Congliang(Beijing Customs Dis-trict,Beijing 100026,China;Zunhua Agricultural and Rural Bureau;Xiamen Customs District;Chinese Academy of Inspection and Quarantine)

机构地区：[1]北京海关,北京100026 [2]遵化市农业农村局 [3]厦门海关 [4]中国检验检疫科学研究院

出　　处：《植物检疫》2021年第2期49-52,共4页Plant Quarantine

基　　金：国家重点研发计划(2016YFF0203203)。

摘　　要：为建立一种快速、灵敏、特异的定量检测南芥菜花叶病毒(arabis mosaic virus,ArMV)的微滴数字RT-PCR方法,根据ArMV外壳蛋白基因保守序列设计了特异性引物和探针,并对该方法的特异性、灵敏度和重复性进行评估。结果表明,该数字RT-PCR检测方法的特异性好,与其他对照病毒均无交叉反应;检测灵敏度可达0.56 copies/μL;3次重复试验变异系数为0.74%,重复性良好。本实验建立的方法适用于实际样品中ArMV的快速定量检测。To develop a rapid,sensitive and specific droplet digital RT-PCR assay for quantitative detection of arabis mosaic virus(ArMV),the primers and probe were designed based on the conserved sequence of ArMV coat protein gene.The specificity,sensitivity and reproducibility of the developed method for ArMV detection were determined.The results showed that the specificity of the developed method was high and there was no crossing reaction with control viruses.The sensitivity of the method was 0.56 copies/μL.The coefficient of variation was 0.74%,indicating the established digital RT-PCR method had good reproducibility.The method developed in this study was suitable for rapid and quantitative detecting ArMV in practical samples.

关 键 词：南芥菜花叶病毒 微滴数字RT-PCR 实时荧光RT-PCR 检测 

分 类 号：S432.41[农业科学—植物病理学]