Department of Microbiology and Immunology, University of British Columbia, Vancouver, British Columbia, Canada.
BugSeq Bioinformatics Inc., Vancouver, British Columbia, Canada.
Clin Microbiol Rev. 2023 Mar 23;36(1):e0011922. doi: 10.1128/cmr.00119-22. Epub 2023 Feb 27.
The advent of next-generation sequencing (NGS) technologies has expanded our ability to detect and analyze microbial genomes and has yielded novel molecular approaches for infectious disease diagnostics. While several targeted multiplex PCR and NGS-based assays have been widely used in public health settings in recent years, these targeted approaches are limited in that they still rely on knowledge of a pathogen's genome, and an untargeted or unknown pathogen will not be detected. Recent public health crises have emphasized the need to prepare for a wide and rapid deployment of an agnostic diagnostic assay at the start of an outbreak to ensure an effective response to emerging viral pathogens. Metagenomic techniques can nonspecifically sequence all detectable nucleic acids in a sample and therefore do not rely on prior knowledge of a pathogen's genome. While this technology has been reviewed for bacterial diagnostics and adopted in research settings for the detection and characterization of viruses, viral metagenomics has yet to be widely deployed as a diagnostic tool in clinical laboratories. In this review, we highlight recent improvements to the performance of metagenomic viral sequencing, the current applications of metagenomic sequencing in clinical laboratories, as well as the challenges that impede the widespread adoption of this technology.
下一代测序(NGS)技术的出现扩展了我们检测和分析微生物基因组的能力,并为传染病诊断带来了新的分子方法。虽然近年来已经有几种靶向多重 PCR 和基于 NGS 的检测方法在公共卫生领域得到广泛应用,但这些靶向方法存在局限性,因为它们仍然依赖于对病原体基因组的了解,而无法检测到未靶向或未知的病原体。最近的公共卫生危机强调了需要在疫情爆发初期准备广泛且快速部署一种无偏倚诊断检测方法,以确保对新出现的病毒病原体做出有效应对。宏基因组技术可以非特异性地对样本中的所有可检测核酸进行测序,因此不依赖于对病原体基因组的事先了解。虽然这项技术已经在细菌诊断方面得到了审查,并在研究中被用于检测和表征病毒,但病毒宏基因组学尚未作为一种诊断工具在临床实验室中得到广泛应用。在这篇综述中,我们重点介绍了宏基因组病毒测序性能的最新改进,以及宏基因组测序在临床实验室中的当前应用,以及阻碍这项技术广泛应用的挑战。
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