CAJ | 학술논문

　　旨在建立可检测马铃薯A病毒(Potato virus A，PVA)的液相芯片快速检测技术，用Primer Premier 5.0软件对GenBank 中 PVA 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PVA的PCR产物杂交反应，用液相芯片检测仪检测荧光信号。结果显示该法具有较好的特异性，不与侵染马铃薯的番茄黑环病毒(Tomato black ring virus，TBRV)、马铃薯纺锤块茎类病毒(Potato spindle tuber viroid，PSTVd)、马铃薯Y病毒(Potato virus Y，PVY)及番茄斑萎病毒(Tomato spotted wilt virus，TSWV)反应；检测灵敏度约为10 pg/μL总RNA。该方法可用于PVA的快速检测。
　　지재건립가검측마령서A병독(Potato virus A，PVA)적액상심편쾌속검측기술，용Primer Premier 5.0연건대GenBank 중 PVA 핵감산서렬진행분석，설계기특이성탐침병용생물소표기。탐침우련형광편마미구후여PVA적PCR산물잡교반응，용액상심편검측의검측형광신호。결과현시해법구유교호적특이성，불여침염마령서적번가흑배병독(Tomato black ring virus，TBRV)、마령서방추괴경류병독(Potato spindle tuber viroid，PSTVd)、마령서Y병독(Potato virus Y，PVY)급번가반위병독(Tomato spotted wilt virus，TSWV)반응；검측령민도약위10 pg/μL총RNA。해방법가용우PVA적쾌속검측。
To develop liquichip method for detecting Potato virus A (PVA), the nucleotide sequence of PVA 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 PVA. Liquichip detection method for PVA was established by using Bio-Rad Liquichip 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 demonstrated better specificity to PVA 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 gel electrophoresis method. The successful detection of PVA in potato sample suggested the feasibility of this procedure for routine testing.