用反转录标记法制备马铃薯病毒的检测芯片  

Detection potato virus by microarray and RT labeling method

作　　者：谷宇[1,3] 杜志游[2] 郎秋蕾[2] 陈集双[2] 何农跃[3] 

机构地区：[1]南京中医药大学,210046 [2]浙江大学微生物分子生物学实验室,杭州 [3]东南大学分子与生物分子电子学教育部重点实验室

出　　处：《植物检疫》2009年第5期5-8,共4页Plant Quarantine

基　　金：江苏省社会公益研究项目(批准号:BM2003701);国家自然科学基金(批准号:60571032)

摘　　要：本文将寡核苷酸芯片和反转录(reversetranscription,RT)荧光标记两种方法相结合,初步建立了芯片检测马铃薯病毒RNA的新方法。针对常见的马铃薯病毒设计了寡核苷酸探针并制备了检测芯片,对从染病植物组织中提取出的RNA进行反转录,同时标记荧光,并进行了多重标记反应。该方法可同时检测4种病毒,虽然检测的灵敏度不够高,但是对于拷贝数较多的植物病毒,反转录标记只需一步反应,不失为一种操作更为简单快速,成本更低的靶序列制备方法,对于检测病毒种类未知的植物样本有一定应用价值。This article establiih a new method to detect potato virus initially. This method combines oligonucleotide microarray technique and fluorescence labeling technique by reverse transcription （RT）. In this research, we designed the specific oligonucleotide probes and fabricate an oligonucleotide microarray, which can detect familiar potato viruses. We labeled the RNA samples extracted from infected potatoes by reverse transcriptase, and tried multiplex RT labeling. After optimizing the protocols, the fabricated gene - chip can detect four kinds of virus at one time. Although the sensitivity is not enough high, this labeling method needs only one - step reaction. So it is a simpler, more rapid and cheaper way to detect plant virus which have rich copies, and can apply to detect plant samples which infect unknown virus.

关 键 词：马铃薯病毒检测 基因芯片 RT标记法 

分 类 号：S435.32[农业科学—农业昆虫与害虫防治]