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. 2018 Aug 21;9:3347. doi: 10.1038/s41467-018-05864-4

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Quantification of NASBA-mediated amplification using toehold switch sensors. a Run-to-run variation in mRNA standards amplified by NASBA and measured by toehold sensors. mRNA standards for the B. thetaiotaomicron species-specific sensor were run in NASBA reactions for 30 min. Outputs from NASBA reactions were used to activate toehold switch sensors in paper-based reactions. b Calibration curve for the B.t. species-specific mRNA. Values from each standard in the individual runs in a were normalized to the 300 fM standard for that specific run and averaged across runs. c Quantifying species-specific mRNAs in stool. E. coli or B. fragilis cells were spiked into 150 mg of a commercial stool sample and processed for total RNA. Species-specific mRNAs were quantified using our paper-based platform and RT-qPCR. d Analysis of clinical stool samples. Six clinical stool samples were processed for total RNA and analyzed by our paper-based platform and RT-qPCR. Data and s.d. are shown in Supplementary Figure 11. e Correlation of clinical sample results. Non-zero paper-based concentrations from d were compared to RT-qPCR determined values. Data represent mean values. Paper-based error bars in a, c, and e represent s.d. from nine replicates (three biological replicates (NASBA reactions) × three technical replicates (paper-based reactions)). RT-qPCR error bars in c and e represent s.d. from six replicates (two biological replicates (RT reactions) × three technical replicates (qPCR reactions))