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. 2021 Feb 1;12:724. doi: 10.1038/s41467-020-20639-6

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© The Author(s) 2021

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. 5 — a Assay sensitivity (using N3 B toehold switch) demonstrated with purified viral RNA. The concentration indicated reflects the viral RNA concentration of the 1 µL aliquot added to NASBA reactions prior to glucose generation. ***p = 0.0001. b The SARS-CoV-2 gene N sensor (N3 B) specificity was tested using viral RNA genomes isolated from influenza A subtypes H1N1 and H7N9, MERS and SARS-CoV-2 using an initial RNA concentration of 1.66 pM. c Sensor N3 B performance upon exposure to RNA extracted from 6 different patient samples (3 SARS-CoV-2 positive and 3 negative patients) assayed with glucose-based sensor. Switch DNA template present at 1.25 ng/µL. Neg: no RNA sample added to NASBA. Cq values from the parallel RT-qPCR assay of these sample listed on the x-axis. ND: not detected. d Companion portable incubator. This device is capable of facilitating the experiment in place of a thermocycler. e Using sensor N3 B, direct comparison between the thermocycler (TC) and portable incubator (PI). SARS-CoV-2 RNA was added to positive (+) reactions at 1.66 pM. For c, statistical analysis is one-way ANOVA. a and b are comprised of 3 technical replicates, representative of 3 independent experiments. e is comprised of 3 biological replicates. ns: not significantly different. Data are presented as mean values ± SD. Source data are provided as a Source Data file.