ZHANG Yong-jiang,XIN Yan-yan,LI Gui-fen,et al.Development of Liquid Chip Technique for Rapid Detection of Tomato Black Ring Virus[J].Northern Horticulture,2013,37(01):127-130.
番茄黑环病毒液相芯片快速检测方法的建立
- Title:
- Development of Liquid Chip Technique for Rapid Detection of Tomato Black Ring Virus
- 文章编号:
- 1001-0009（2013）01-0127-04
- 关键词:
- 番茄黑环病毒（TBRV）; 液相芯片技术; 检测
- 分类号:
- S 436.412.1+1
- 文献标志码:
- A
- 摘要:
- 为建立番茄黑环病毒（Tomato black ring virus，TBRV）的液相芯片检测技术，对GenBank中的TBRV核苷酸序列进行分析，设计其特异性探针并用生物素标记。探针偶联荧光编码微球后与TBRV的RT-PCR产物进行杂交，用液相芯片检测仪检测荧光信号。结果表明：建立的方法具有良好的特异性，能够有效区分侵染马铃薯的TBRV、马铃薯纺锤块茎类病毒（PSTVd）、马铃薯A病毒（PVA）及番茄斑萎病毒（TSWV）；检测灵敏度约为1 pg/μL总RNA，与常规RT-PCR灵敏度相当。实际样品检测结果表明该方法可用于TBRV的日常检测。
- Abstract:
- To develop liquid chip technique for detecting Tomato black ring virus (TBRV)，the nucleotide sequence of TBRV was analyzed by using the software Primer Premier 5.0，and specific probe labeled with biotin was prepared and coupled with fluorescence-coded microspheres，which was used for hybridization reaction to RT-PCR products of TBRV.Liquid chip detection method for TBRV was established by using Bio-Rad liquid chip to detect fluorescence signal in the reaction system.The specificity of this method was evaluated by applying the proposed method to detect four viruses infecting potato，TBRV，Potato spindle tuber viroid (PSTVd)，Potato virus A (PVA) and Tomato spotted wilt virus (TSWV).The results showed that better specificity to TBRV and no reaction with other three viruses.The sensitivity of the method was 1 pg/μL of total RNA，which was equal to the conventional RT-PCR method.The successful detection of TBRV in potato samples suggested the feasibility of this procedure for routine testing.
参考文献/References:
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备注/Memo
第一作者简介：张永江（1977-），男，博士，副研究员，现主要从事植物检疫研究工作。E-mail:zhangyjpvi@yeah.net.