Institute for Medical Engineering and Science, Massachusetts Institute of Technology, 77 Massachusetts Ave, Cambridge, MA, 02139, USA.
Division of Gastroenterology, Massachusetts General Hospital, 55 Fruit Street, Boston, MA, 02114, USA.
Nat Commun. 2018 Aug 21;9(1):3347. doi: 10.1038/s41467-018-05864-4.
There is a need for large-scale, longitudinal studies to determine the mechanisms by which the gut microbiome and its interactions with the host affect human health and disease. Current methods for profiling the microbiome typically utilize next-generation sequencing applications that are expensive, slow, and complex. Here, we present a synthetic biology platform for affordable, on-demand, and simple analysis of microbiome samples using RNA toehold switch sensors in paper-based, cell-free reactions. We demonstrate species-specific detection of mRNAs from 10 different bacteria that affect human health and four clinically relevant host biomarkers. We develop a method to quantify mRNA using our toehold sensors and validate our platform on clinical stool samples by comparison to RT-qPCR. We further highlight the potential clinical utility of the platform by showing that it can be used to rapidly and inexpensively detect toxin mRNA in the diagnosis of Clostridium difficile infections.
需要开展大规模的纵向研究，以确定肠道微生物组及其与宿主相互作用影响人类健康和疾病的机制。目前用于分析微生物组的方法通常采用下一代测序技术，这种方法昂贵、缓慢且复杂。在这里，我们提出了一种使用基于纸张的无细胞反应中的 RNA 触发开关传感器来进行微生物组样本的经济实惠、按需且简单分析的合成生物学平台。我们证明了针对影响人类健康的 10 种不同细菌和 4 种临床相关宿主生物标志物的 mRNA 的物种特异性检测。我们开发了一种使用我们的触发传感器定量 mRNA 的方法，并通过与 RT-qPCR 的比较，在临床粪便样本上验证了我们的平台。我们通过展示该平台可用于快速且经济地检测艰难梭菌感染诊断中的毒素 mRNA，进一步强调了该平台的潜在临床应用价值。
