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. 2018 Aug 13;9:3238. doi: 10.1038/s41467-018-05733-0

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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. 3 — Programmability of enVision. a Schematic of the programmable recognition nanostructure. The hybrid structure consists of a conserved sequence region, that binds to inactivate DNA polymerase (pol), and a variable region (duplex and overhang segments) that can be made complementary to target DNA. Not drawn to scale. b Effects of target mismatches in the variable region. Synthetic DNA targets, designed to have varying numbers of mismatches against the variable region, were incubated with the recognition nanostructure. All signals were normalized against that of the complementary DNA target (0 mismatch). Mismatches against the duplex region produced significantly lower signals (*P < 0.05, ** P < 0.005, ***P < 0.0005, n.s. not significant, Student’s t test). c Pan-HPV recognition. Two pan-HPV recognition nanostructures were developed according to the HPV consensus genome, to harbor different numbers of mismatches against DNA targets obtained from six HPV subtypes. All mismatches were mapped to the duplex and overhang regions. Nanostructure 1, that accommodated more mismatches in the overhang region, demonstrated better pan-recognition capability. All signals were normalized against that of the complementary DNA target (positive). d Comparison of enVision and qPCR measurements for specific HPV subtyping. At a cutoff for 100% sensitivity, enVision had 100% specificity (56/56) while SYBR Green-based qPCR had 92.9% specificity (52/56). Specific nanostructures were designed according to a highly variable region of the HPV genome with sequence variations contained within the sensitive duplex region. DNA targets from different HPV subtypes were measured via color intensity through the enVision smartphone platform (left) and cycle counts through the SYBR-Green qPCR system (right). All signals were acquired relative to appropriate controls (i.e., water as a no-template control). Signals from respective detection systems were globally presented in the form of heat maps for comparisons of assay performance. All measurements were performed in triplicate, and the data are displayed as mean ± s.d. in b and c