Figure 6

LAMP in FLIPP-NAAT from dry-stored reagents. (A) End-point imaging of the FLIPP-NAAT device after 5 minutes of addition of SYBR green I dye. Numbers on top of each reaction zone indicate log₁₀ of starting copy number. N – no template control. (B) Mean pixel intensity of the green color from all starting copy numbers was statistically higher (*P < 0.05; N = 3) from that of no template control. 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. (C) LAMP in FLIPP-NAAT from dry-stored reagents and rehydration with mock sputum. Each reaction zone had 100 copies of Mtb gDNA as the starting template. The markings on top of each of the reaction zone indicate the dilution of mock sputum used for rehydration. 3 W, 2 W, 1 W – reaction set-up with template suspended in water containing 1000, 100 and 10 copies of Mtb gDNA respectively, as positive controls. All reactions worked despite the presence of the complex sputum matrix.