Reverse Transcription Lesion-Induced DNA Amplification: An Instrument-Free Isothermal Method to Detect RNA

27 April 2020, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

One challenge in point-of-care diagnostics is the lack of room-temperature methods for RNA detection based on enzymatic amplification and visualization steps. Here we perform a reverse transcription ligase chain reaction using our isothermal lesion induced DNA amplification (LIDA) technique that can be tuned to operate at any desired temperature. Using RNA-triggered LIDA, we can detect as little as ~100 attomoles target RNA and can distinguish RNA target from total cellular RNA. Finally, we demonstrate that the resulting DNA amplicons can be detected colorimetrically, also at room temperature, by rapid, target-triggered disassembly of DNA-modified gold nanoparticles. This integrated amplification/detection platform requires no heating or visualization instrumentation, which is an important step towards realizing instrument-free POC testing.

Keywords

Lesion-induced DNA amplification
DNA modified gold nanoparticles
RNA detection
Colorimetric method
Room temperature amplification and detection

Supplementary materials

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