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US 11,952,637 B2Granted
Rapid low-cost detection of SARS-CoV-2 using isothermal amplification and sensing methods
AssigneesArizona Board of Regents on behalf of Arizona State University, The Governing Council of the University of Toronto
InventorsAlexander Green, Keith Pardee, Margot Karlikow, Kaiyue Wu, Masoud Norouzi
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Abstract
Provided herein are methods and compositions for rapid, highly sensitive detection of SARS-CoV-2 in biological samples. In particular, provided herein is a rapid, low-cost method for detecting SARS-CoV-2 that provides reliable, visible test results and does not require PCR reagents, elaborate biosafety precautions, or sophisticated laboratory equipment.
Claims19
A method of detecting a viral SARS-CoV-2 RNA in a sample from a subject, the method comprising the steps of:
- (a) heating the sample, whereby viral RNAs are released from viral particles present in the sample;
- (b) amplifying the released viral RNAs, wherein amplifying comprises isothermal amplification and produces a target nucleic acid;
- and
- (c) contacting the amplified target nucleic acid to one or more riboregulators, wherein each riboregulator comprises:
- (i) a first region that is complementary to at least a portion of the target nucleic acid or a reverse complement thereof;
- (ii) a second region that encodes a reporter protein and translation regulatory sequences;
- wherein the contacting occurs under conditions that allow translation of the reporter protein in the presence of the target nucleic acid or reverse complement thereof, but not in the absence of the target nucleic acid or reverse complement thereof;
- and
- (d) detecting the reporter protein as an indicator that the viral SARS-CoV-2 RNA is present in the subject sample;
- wherein each riboregulator first region comprises, independently, one of SEQ ID NOs:
- 26-51.
8 more elements
Dependencies are inferred from the claim text.