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用于快速鉴定植物病毒的微阵列。

Microarrays for rapid identification of plant viruses.

作者信息

Boonham Neil, Tomlinson Jenny, Mumford Rick

机构信息

Central Science Laboratory, Sand Hutton, York, YO41 1LZ, United Kingdom.

出版信息

Annu Rev Phytopathol. 2007;45:307-28. doi: 10.1146/annurev.phyto.45.062806.094349.

Abstract

Many factors affect the development and application of diagnostic techniques. Plant viruses are an inherently diverse group that, unlike cellular pathogens, possess no nucleotide sequence type (e.g., ribosomal RNA sequences) in common. Detection of plant viruses is becoming more challenging as globalization of trade, particularly in ornamentals, and the potential effects of climate change enhance the movement of viruses and their vectors, transforming the diagnostic landscape. Techniques for assessing seed, other propagation materials and field samples for the presence of specific viruses include biological indexing, electron microscopy, antibody-based detection, including enzyme-linked immunosorbent assay (ELISA), polymerase chain reaction (PCR), and microarray detection. Of these, microarray detection provides the greatest capability for parallel yet specific testing, and can be used to detect individual, or combinations of viruses and, using current approaches, to do so with a sensitivity comparable to ELISA. Methods based on PCR provide the greatest sensitivity among the listed techniques but are limited in parallel detection capability even in "multiplexed" applications. Various aspects of microarray technology, including probe development, array fabrication, assay target preparation, hybridization, washing, scanning, and interpretation are presented and discussed, for both current and developing technology.

摘要

许多因素影响着诊断技术的发展与应用。植物病毒本质上是一个多样化的群体,与细胞病原体不同,它们没有共同的核苷酸序列类型(例如核糖体RNA序列)。随着贸易全球化,尤其是观赏植物贸易的全球化,以及气候变化的潜在影响促进了病毒及其传播媒介的移动,改变了诊断格局,植物病毒的检测变得更具挑战性。评估种子、其他繁殖材料和田间样本中特定病毒存在情况的技术包括生物索引、电子显微镜、基于抗体的检测(包括酶联免疫吸附测定法(ELISA))、聚合酶链反应(PCR)和微阵列检测。其中,微阵列检测提供了最大的平行且特异性检测能力,可用于检测单个病毒或病毒组合,并且按照目前的方法,其检测灵敏度可与ELISA相媲美。在所列技术中,基于PCR的方法灵敏度最高,但即使在“多重”应用中,其平行检测能力也有限。本文针对当前技术和正在发展的技术,介绍并讨论了微阵列技术的各个方面,包括探针开发、芯片制作、检测靶标制备、杂交、洗涤、扫描和解读。

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