Fig 3. Detection limit of RT-LAMP and RT-PCR assay using total RNA extracts.
Extracted total RNA from a P. falciparum-infected blood sample was serially diluted (103to 10-3ng/μL) and 2.5μL tested by both RT-LAMP and RT-PCR methods. A) Agarose (1.2%) gel analyses and visualization of DNA laddering in RT-LAMP amplified samples. B) Agarose (1.2%) gel analyses of the PfExp1 RT-PCR products, showing similarities in the detection limits of both RT-LAMP and RT-PCR methods. Mk: 1Kb DNA ladder, NTC: no-template control reaction tube, and numbers on each lane indicate the total RNA concentration (in ng/μL) in each tube prior to the assays.C) Linear regression analyses showing a significant inverse correlation between total RNA amounts and the time-to-peak amplification signal for the RT-LAMP method. Data is representative ofthe lowest limit of detection obtained from three replicate experiments at different diluted RNA concentration ranges.