对来自高度异质样本的单一病毒物种进行全基因组测序。
Whole-Genome Sequencing of a Single Viral Species from a Highly Heterogeneous Sample.
作者信息
Han Hee-Sun, Cantalupo Paul G, Rotem Assaf, Cockrell Shelley K, Carbonnaux Martial, Pipas James M, Weitz David A
机构信息
Department of Physics, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA 02138 (USA).
Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA 15260 (USA).
出版信息
Angew Chem Int Ed Engl. 2015 Nov 16;54(47):13985-8. doi: 10.1002/anie.201507047. Epub 2015 Aug 28.
Abstract
Metagenomic studies suggest that only a small fraction of the viruses that exist in nature have been identified and studied. Characterization of unknown viral genomes is hindered by the many genomes populating any virus sample. A new method is reported that integrates drop-based microfluidics and computational analysis to enable the purification of any single viral species from a complex mixed virus sample and the retrieval of complete genome sequences. By using this platform, the genome sequence of a 5243 bp dsDNA virus that was spiked into wastewater was retrieved with greater than 96% sequence coverage and more than 99.8% sequence identity. This method holds great potential for virus discovery since it allows enrichment and sequencing of previously undescribed viruses as well as known viruses.
摘要
宏基因组学研究表明，自然界中存在的病毒只有一小部分已被鉴定和研究。任何病毒样本中存在的众多基因组阻碍了对未知病毒基因组的表征。据报道，一种新方法将基于液滴的微流控技术与计算分析相结合，能够从复杂的混合病毒样本中纯化出任何单一病毒种类，并获取完整的基因组序列。通过使用该平台，从废水中添加的一种5243 bp双链DNA病毒的基因组序列被检索到，序列覆盖率大于96%，序列同一性超过99.8%。这种方法在病毒发现方面具有巨大潜力，因为它允许对以前未描述的病毒以及已知病毒进行富集和测序。
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