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. 2019 Feb 11;9:1739. doi: 10.1038/s41598-018-38056-7

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© The Author(s) 2019

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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Detection of V. cholerae amplification using purified DNA. NTC represents no added V. cholerae DNA. (A) Real-time fluorescence was monitored over a 20-minute LAMP reaction for initial DNA concentrations between 10⁰–10⁵ DNA copies/reaction and (B) the corresponding C_T values were recorded for each reaction and are not available (NA) for samples that did not amplify. (C) A 2% agarose gel confirms amplification and presents the DNA banding pattern of LAMP amplicons at the different dilutions. (D) Box plots of the average change in fluorescence ( $Δ E v a G r e e n / R o x$ ) shows a trend of a greater change in fluorescence signal at higher initial V. cholerae DNA concentrations with statistical differences for samples 10² (***p-value < 0.001), 10³, 10⁴, and 10⁵ (****p-value < 0.0001) DNA copies/reaction when compared to NTC. (E) Particle diffusometry measurements of the viscosity change of LAMP reactions show statistically significant measurements for 10² (*p-value < 0.05), 10⁴, and 10⁵ (****p-value < 0.0001) DNA copies/reaction when compared to NTC. (D,E) Statistical analysis was a one-way ANOVA with Dunnett’s post-hoc relative to NTC (n = 3). (F) A positive correlation between change in fluorescence and PD yields a Pearson’s correlation coefficient of 0.81.