Viral discovery and sequence recovery using DNA microarrays
Viral discovery and sequence recovery using DNA microarrays
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DOI：
10.1371/journal.pbio.0000002
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发表时间：
2003-11-01
期刊：
影响因子：
9.8
通讯作者：
DeRisi, JL
中科院分区：
文献类型：
--
作者：
Wang, D;Urisman, A;DeRisi, JL
由于新出现的传染病带来的持续威胁以及用于鉴定新病原体的现有方法的局限性，对用于病毒发现的新技术方法存在巨大需求。我们在此描述了一种基于DNA微阵列的新型病毒鉴定和表征平台。这种方法的核心是一种DNA微阵列，旨在检测广泛的已知病毒以及现有病毒家族的新成员;这种微阵列包含来自GenBank中每个完全测序的参考病毒基因组的最高度保守的70聚体序列。在2003年3月严重急性呼吸综合征（SARS）爆发期间，与该微阵列的杂交揭示了从SARS患者培养的病毒分离物中存在以前未表征的冠状病毒。为了进一步表征这种新病毒，通过物理回收与单个阵列元件杂交的病毒序列来克隆约1 kb的未知病毒基因组。对这些片段的测序证实，该病毒确实是冠状病毒家族的新成员。这种阵列杂交与直接病毒序列恢复的组合应被证明是快速鉴定和表征新型病毒和新兴传染病的一般策略。
Because of the constant threat posed by emerging infectious diseases and the limitations of existing approaches used to identify new pathogens, there is a great demand for new technological methods for viral discovery. We describe herein a DNA microarray-based platform for novel virus identification and characterization. Central to this approach was a DNA microarray designed to detect a wide range of known viruses as well as novel members of existing viral families; this microarray contained the most highly conserved 70mer sequences from every fully sequenced reference viral genome in GenBank. During an outbreak of severe acute respiratory syndrome (SARS) in March 2003, hybridization to this microarray revealed the presence of a previously uncharacterized coronavirus in a viral isolate cultivated from a SARS patient. To further characterize this new virus, approximately 1 kb of the unknown virus genome was cloned by physically recovering viral sequences hybridized to individual array elements. Sequencing of these fragments confirmed that the virus was indeed a new member of the coronavirus family. This combination of array hybridization followed by direct viral sequence recovery should prove to be a general strategy for the rapid identification and characterization of novel viruses and emerging infectious disease.