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. 2019 Oct 25;9:15367. doi: 10.1038/s41598-019-51873-8

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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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LAMP in FLIPP-NAAT and end-point fluorescence detection. (A) Testing uniformity of field in FLIPP-NAAT. Numbers on each reaction zone mark the 12 replicates. The mean intensity for signals from the 12 reaction zones was 160 with a coefficient of variation of only 5.4%. (B) Cell phone image of FLIPP-NAAT after 5 minutes of addition of SYBR Green I dye for fluorescence detection. Numbers on top of each reaction zone indicate log₁₀ of Mtb gDNA starting copy number. N – no template control. (C) Mean pixel intensity of green color for the different starting copy numbers. The mean intensity of each starting copy number was statistically different from that of others (*P < 0.05; N = 3). (D) LAMP reactions in FLIPP-NAAT in presence of 3.1 * 10³ copies of human gDNA. Numbers on top of each reaction zone indicate log₁₀ of Mtb gDNA starting copy number. N – no template control containing only human gDNA. (E) Mean pixel intensity of green color for the different starting copy numbers. The mean intensity of 100 and 10 copy number was statistically different from that of the negative controls (*P < 0.05; N = 3). The one drop out for 10 copies (second reaction zone from the right in the bottom row, Panel D) was not included for the intensity analysis as it was a false negative. The dashed line represents the threshold value of (µ_N + 3σ_N). Samples with intensity below the threshold were considered as TB negative while samples with green color intensity above the threshold were considered as TB positive.