双重DPO-RT-PCR检测南芥菜花叶病毒及菜豆荚斑驳病毒  

Dual priming oligonucleotide system for the detection of Arabis mosaic virus and Bean pod mottle virus

作　　者：袁俊杰 马新华 谢小燕 卢乃会 杨卓瑜 陈文 何群 赵玲 龙阳 Yuan Junjie;Ma Xinhua;Xie Xiaoyan;Lu Naihui;Yang Zhuoyu;Chen Wen;He Qun;Zhao Ling;Long Yang(Zhanjiang Customs,Zhanjiang Guangdong 524000,China;Zengcheng Customs,Guangzhou Guangdong 510000,China)

机构地区：[1]湛江海关,广东湛江524000 [2]增城海关,广东广州510000

出　　处：《中国植保导刊》2021年第6期92-96,共5页China Plant Protection

基　　金：国家重点研发计划(2018YFC0809200);海关总署科研项目(2019HK052)。

摘　　要：针对南芥菜花叶病毒(Arabis mosaic virus,Ar MV)及菜豆荚斑驳病毒(Bean pod mottle virus,BPMV)基因组中特异性较高的序列,分别设计了特异性DPO(dual priming oligonucleotide)引物,建立了一种快速检测这两种植物病毒的多重DPO-RT-PCR方法,并评价其特异性、灵敏度及退火温度敏感性。设计的DPO引物特异性较强,Ar MV和BPMV可分别扩增出217 bp与800 bp的特异性条带,检测灵敏度均达0.02 ng/μL。该检测方法对退火温度不敏感。实际样本及模拟样本的检测结果均与国家标准方法一致。本研究建立的检测方法可应用于Ar MV、BPMV的快速筛查,为口岸检疫监管提供技术支撑。In this study,two pairs of DPO(dual priming oligonucleotide)primers were designed based on the specific sequence of Ar MV and BPMV,and a multiplex DPO-PCR method had been developed for the detection of Ar MV and BPMV.The specificity and sensitivity of the method had been tested.The result showed that the DPO primers were highly sequence-specific,with which the 217 bp amplified fragment of Ar MV and 800 bp fragment of BPMV could be obtained,and the sensitivity of the method was 0.02 ng/μL for the two viruses.This method was not sensitive to annealing temperature.The test results of this method for the actual samples and the simulant were consistent with that of the national standard.The multiplex DPO-PCR method established in this study can be used for rapid screening for Ar MV and BPMV,and provide technical support for the quarantine inspection at ports.

关 键 词：南芥菜花叶病毒 菜豆荚斑驳病毒 DPO-RT-PCR 

分 类 号：S41-31[农业科学—植物保护]