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Streaming algorithms for identification of pathogens and antibiotic resistance potential from real-time MinIONTM sequencing

机译:通过实时MinIONTM测序来识别病原体和抗生素耐药性的流算法

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摘要

The recently introduced Oxford Nanopore MinION platform generates DNA sequence data in real-time. This has great potential to shorten the sample-to-results time and is likely to have benefits such as rapid diagnosis of bacterial infection and identification of drug resistance. However, there are few tools available for streaming analysis of real-time sequencing data. Here, we present a framework for streaming analysis of MinION real-time sequence data, together with probabilistic streaming algorithms for species typing, strain typing and antibiotic resistance profile identification. Using four culture isolate samples, as well as a mixed-species sample, we demonstrate that bacterial species and strain information can be obtained within 30 min of sequencing and using about 500 reads, initial drug-resistance profiles within two hours, and complete resistance profiles within 10 h. While strain identification with multi-locus sequence typing required more than 15x coverage to generate confident assignments, our novel gene-presence typing could detect the presence of a known strain with 0.5x coverage. We also show that our pipeline can process over 100 times more data than the current throughput of the MinION on a desktop computer.Electronic supplementary materialThe online version of this article (doi:10.1186/s13742-016-0137-2) contains supplementary material, which is available to authorized users.
机译:最近推出的牛津纳米孔MinION平台可实时生成DNA序列数据。这具有缩短样品到结果时间的巨大潜力,并可能具有诸如快速诊断细菌感染和鉴定耐药性等优点。但是,很少有工具可用于实时测序数据的流分析。在这里,我们介绍了MinION实时序列数据流分析的框架,以及用于物种类型,菌株类型和抗生素抗性概况鉴定的概率流算法。使用四个分离培养物样品以及一个混合物种样品,我们证明可以在测序的30分钟内获得细菌种类和菌株信息,并使用约500个读数,两个小时内的初始耐药性谱以及完整的耐药性谱10小时内。虽然使用多位点序列分型鉴定菌株需要超过15倍的覆盖率才能产生可​​信的分配,但我们的新型基因存在分型可以检测到覆盖度为0.5倍的已知菌株的存在。我们还表明,与台式机上的MinION当前的吞吐量相比,我们的管道可以处理的数据多100倍。电子补充材料本文的在线版本(doi:10.1186 / s13742-016-0137-2)包含补充材料,可供授权用户使用。

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