Development and Validation of Rapid Colorimetric Reverse Transcription Loop-Mediated Isothermal Amplification for Detection of Rift Valley Fever Virus

Figure 9

Detection limit of the new colorimetric-RVFV-UDG-RT-LAMP assay. (a) Analytical sensitivity of qRT-PCR of six replicates determined by tenfold dilution of known concentration of RVFV total RNA (18.5 pg–1850000 pg). (b) Standard curve generated by plotting the number of RVFV RNA copies (x-axis), and the mean of the corresponding qRT–PCR threshold cycle (Ct) of three independent experiments on the y-axis. (c) Limit of detection of the RVFV-UDG-RT-LAMP assay using a tenfold dilution of known concentration of RVFV total RNA as described under qRT-PCR as monitored through color change and resolved on 1% agarose gel electrophoresis. The M-100 bp molecular ladder (Cat SM0241, Thermo Fisher Scientific Inc., USA), 7-NTC, 1–6 tenfold dilution from 18.5 pg to 1.85 × 10⁶, respectively.

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