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

preprint
submitted on 24.04.2020 and posted on 27.04.2020 by Bibi Safeenaz Alladin-Mustan, Jesse Yuzik, Daria Raquel Queiroz de Almeida, Yuning Liu, Yimeng Li, Camilla Mendes, Julianne Gibbs

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.

Funding

Alfred P. Sloan Foundation for a Research Fellowship

Future Energy Systems

Natural Sciences and Engineering Research Council of Canada (NSERC): Accelerator Award

Alberta Innovates Technology Futures (AITF)

Alberta/Technical University of Munich Graduate School for Hybrid Functional Materials (ATUMS-NSERC CREATE)

History

Email Address of Submitting Author

gibbsdav@ualberta.ca

Institution

University of Alberta

Country

Canada

ORCID For Submitting Author

0000-0001-5819-2306

Declaration of Conflict of Interest

No conflict of interest.

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