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. Author manuscript; available in PMC: 2019 Mar 13.
Published in final edited form as: J Vis Exp. 2018 Mar 13;(133):10.3791/57051. doi: 10.3791/57051

Figure 3. RT-LAMP in real-time.

Figure 3

(A) Singleplex and multiplexed limit of detection (LOD) for Zika (ZV) using 80 nM FAM-labeled strand-displacing probe in real-time using real-time PCR instrument (channel 483-533 nm). Samples were incubated at 65°C for about 50 min. (B) Fluorescence was observed though an orange filter followed by the excitation of the samples at 470 nm with blue light. Samples from A were used for visualization. (C) Real-time analysis for LOD on Chikungunya (CH) target using 80 nM of HEX-bearing LAMP probes in singleplex format using fluorescence channel of 523-568 nm on a real-time PCR instrument. (D) Real-time analysis for LOD on Dengue 1 (D1) targets using 80 nM of TAMRA-bearing LAMP probes in singleplex format using fluorescence channel of 558-610 nm on a real-time PCR instrument. (E) Determination of maximum urine content in RT-LAMP reaction. A range of final urine concentrations (0 %-50 %) were tested using 80 nM FAM-labeled probe in the presence of 2.85 pfu of Zika vRNA. NTC = No template control on all panels. Reprinted with permission from Yaren et al. 201720.