Department of Medicine and.
Department of Physiology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
J Clin Invest. 2022 Apr 1;132(7). doi: 10.1172/JCI154944.
Next-generation sequencing (NGS) technology has advanced our understanding of the human microbiome by allowing for the discovery and characterization of unculturable microbes with prediction of their function. Key NGS methods include 16S rRNA gene sequencing, shotgun metagenomic sequencing, and RNA sequencing. The choice of which NGS methodology to pursue for a given purpose is often unclear for clinicians and researchers. In this Review, we describe the fundamentals of NGS, with a focus on 16S rRNA and shotgun metagenomic sequencing. We also discuss pros and cons of each methodology as well as important concepts in data variability, study design, and clinical metadata collection. We further present examples of how NGS studies of the human microbiome have advanced our understanding of human disease pathophysiology across diverse clinical contexts, including the development of diagnostics and therapeutics. Finally, we share insights as to how NGS might further be integrated into and advance microbiome research and clinical care in the coming years.
下一代测序（NGS）技术通过发现和描述无法培养的微生物，并预测其功能，使我们对人类微生物组有了更深入的了解。关键的 NGS 方法包括 16S rRNA 基因测序、宏基因组测序和 RNA 测序。对于临床医生和研究人员来说，为特定目的选择哪种 NGS 方法通常并不清楚。在这篇综述中，我们描述了 NGS 的基本原理，重点介绍了 16S rRNA 和宏基因组测序。我们还讨论了每种方法的优缺点，以及数据变异性、研究设计和临床元数据收集方面的重要概念。我们进一步展示了 NGS 研究人类微生物组如何在不同的临床环境中推进我们对人类疾病病理生理学的理解的例子，包括诊断和治疗方法的发展。最后，我们分享了关于 NGS 如何在未来几年进一步整合到微生物组研究和临床护理中并推动其发展的见解。
