Reverse transcription lesion-induced DNA amplification: An instrument-free isothermal method to detect RNA

One challenge in point-of-care (POC) diagnostics is the lack of room-temperature methods for RNA detection based on enzymatic amplification and visualization steps. Here we perform reverse transcription lesion-induced DNA (RT-LIDA), an isothermal method that only requires T4 ligase. RT-LIDA involves RNA-templated ligation primers to form complementary (cDNA) followed by toehold-mediated strand displacement cDNA its exponential via our ligase chain reaction LIDA. Each step tuned proceed at 28 °C, which falls within range global room temperatures. Using RT-LIDA, can detect as little ?100 amol target distinguish from total cellular RNA. Finally, demonstrate resulting amplicons be detected colorimetrically, also temperature, rapid, target-triggered disassembly DNA-modified gold nanoparticles. This integrated amplification/detection platform no heating or instrumentation, important towards realizing instrument-free POC testing.