Detection of potato viruses using microarray technology: towards a generic method for plant viral disease diagnosis
Detection of potato viruses using microarray technology: towards a generic method for plant viral disease diagnosis
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DOI：
10.1016/s0166-0934(02)00284-7
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发表时间：
2003-03-01
影响因子：
3.1
通讯作者：
Barker, I
中科院分区：
文献类型：
--
作者：
Boonham, N;Walsh, K;Barker, I
目前，大多数诊断方法都是针对检测非常特定的目标物种，并且通常需要并行运行多个测定以获得结果。可用于病毒检测的通用方法倾向于仅对属水平进行鉴定。本文中描述的方法通过利用具有在单个测定中测试大量靶标的潜力的技术来解决这个问题。利用所构建的芯片，该方法能够检测几种常见的马铃薯病毒（PVY，PVX，PVA，PVS）的单一和混合感染。该方法显示能够区分具有小于80%序列同一性的序列，但能够检测具有大于90%序列同一性的序列变体。因此，该方法应该是有用的区分在物种水平上，但能够很好地科普内发现的RNA病毒基因组的内在变异性。发现该测定的灵敏度与ELISA相当。这篇论文通过首次提出一种理论上不仅可以用于病毒，而且可以用于现代诊断实验室想要测试的所有植物病原体和害虫的方法，以一种完全通用和高度并行的格式，说明了诊断方法学发展的重要一步。(C)2002 Elsevier Science B.V.保留所有权利。
Currently, most diagnostic methodology is geared towards detection of a very specific target species and often a number of assays need to be run in parallel to reach a result. The generic methods that are available for virus testing tends to give identification to the genus level only. The method described in this paper addresses this problem by exploiting a technology that has potential to test for a large number of targets in a single assay. Using the array constructed, the method was able to detect several common potato viruses (PVY, PVX, PVA, PVS) in single and mixed infections. The method was shown to be able to discriminate sequences with less than 80% sequence identity but was able to detect sequence variants with greater than 90% sequence identity. Thus the method should be useful for discriminating at the species level, but able to cope well with the intrinsic variability found within the genomes of RNA viruses. The sensitivity of the assay was found to be comparable with ELISA. The paper illustrates a significant step forward in the development of diagnostic methodologies by presenting for the first time a method that could theoretically be used not just for viruses, but for all the plant pathogens and pests that a modern diagnostic laboratory would want to test for, in a single completely generic and highly parallel format. (C) 2002 Elsevier Science B.V. All rights reserved.