大麦条纹花叶病毒实时荧光RT-PCR检测方法的建立及应用  

Establishment and application of a real-time RT-PCR detection method for Barley Stripe Mosaic Virus

作　　者：王佳莹 徐瑛 李文[2] 崔俊霞 张吉红 陈先锋 Wang Jiaying;Xu Ying;Li Wen;Cui Junxia;Zhang Jihong;Chen Xianfeng(Ningbo Academy of Inspection and Quarantine,Ningbo 315012,China;Ningbo City College of Vocational Technology)

机构地区：[1]宁波检验检疫科学技术研究院,浙江宁波315012 [2]宁波城市职业技术学院

出　　处：《植物检疫》2019年第1期44-47,共4页Plant Quarantine

基　　金：宁波市科技创新团队项目(2015C110018);宁波市自然科学基金(2018A610215)

摘　　要：大麦条纹花叶病毒为禾本科作物上一种重要的病原,可造成严重的经济损失。该病毒的主要传播方式为种传,因此对种子进行检疫监管是控制病害传播的一种有效途径。本文依据病毒保守序列RNA2beta C蛋白设计引物,研发了一种实时荧光RT-PCR检测方法。经特异性与灵敏度评价,仅对大麦条纹花叶病毒有效,无法对小麦线条花叶病毒、甘蔗花叶病毒及玉米褪绿斑驳病毒的阳性样品进行鉴定;且扩增灵敏度与普通RT-PCR相当,处于0.02 ng/μL水平。该方法相较于已有的检测方法,操作更简便快速,不易污染,灵敏度高,特异性优。此外,本研究结合实时荧光RT-PCR与普通RT-PCR两种检测方法,对未知谷物粉进行大麦条纹花叶病毒的检测鉴定,证实了新方法的可行性。Barley stripe mosaic virus is an important pathogen on gramineous crops,which can cause considerable economic loss.BSMV spreads via seeds mainly.Accordingly,seed quarantine is an effective way to control virus spread.In this paper,one set of primers was designed based on the conservative sequence in RNA2 beta C protein and a real-time RT-PCR method was established for BSMV quarantine. Specificity and sensitivity showed that this method could be only used for BSMV,but not Wheat streak mosaic virus,Sugarcane mosaic virus or Maize chlorotic mottle virus.Sensitivity test confirmed that this method detected samples at the level of 0.02 ng/μL,similar to normal RT-PCR.The real-time RT-PCR established in this study is simpler,faster,less contaminative,more sensitive and more specific compared to other existing methods.What's more,this method was combined with normal RT-PCR in BSMV detection and identification of an unknown crop powder sample ,which turned out to be highly feasible.

关 键 词：大麦条纹花叶病毒 实时荧光RT-PCR 灵敏度 特异性 快速筛查 

分 类 号：S41[农业科学—植物保护]