TY - JOUR T1 - Engineered RNA biosensors enable ultrasensitive SARS-CoV-2 detection in a simple color and luminescence assay. JF - Life Sci Alliance Y1 - 2021 A1 - Chakravarthy, Anirudh A1 - Nandakumar, Anirudh A1 - George, Geen A1 - Ranganathan, Shyamsundar A1 - Umashankar, Suchitta A1 - Shettigar, Nishan A1 - Palakodeti, Dasaradhi A1 - Gulyani, Akash A1 - Ramesh, Arati KW - Biosensing Techniques KW - COVID-19 KW - Humans KW - Luminescence KW - Nucleic Acid Amplification Techniques KW - RNA KW - RNA, Viral KW - SARS-CoV-2 AB -

The continued resurgence of the COVID-19 pandemic with multiple variants underlines the need for diagnostics that are adaptable to the virus. We have developed toehold RNA-based sensors across the SARS-CoV-2 genome for direct and ultrasensitive detection of the virus and its prominent variants. Here, isothermal amplification of a fragment of SARS-CoV-2 RNA coupled with activation of our biosensors leads to a conformational switch in the sensor. This leads to translation of a reporter protein, for example, LacZ or nano-lantern that is easily detected using color/luminescence. By optimizing RNA amplification and biosensor design, we have generated a highly sensitive diagnostic assay that is capable of detecting as low as 100 copies of viral RNA with development of bright color. This is easily visualized by the human eye and quantifiable using spectrophotometry. Finally, this PHAsed NASBA-Translation Optical Method (PHANTOM) using our engineered RNA biosensors efficiently detects viral RNA in patient samples. This work presents a powerful and universally accessible strategy for detecting COVID-19 and variants. This strategy is adaptable to further viral evolution and brings RNA bioengineering center-stage.

VL - 4 IS - 12 ER -