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US 2023/0416849 A1Published application
Methods for detecting viral rna using atp-releasing nucleotides
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsEric T. Kool, Michael G. Mohsen
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Abstract
Compositions and methods are provided for a biochemical method for detecting virus RNA sequences, either after isolation or directly in patient samples, by detecting ATP released from an ATP-releasing nucleotide (ARN) during synthesis. In some embodiments the virus is a coronavirus, e.g. SARS-CoV2.
Claims14
A method for detecting the presence of a virus RNA sequence in a sample comprising nucleic acids, the method comprising:
- contacting the nucleic acid with a reaction mixture comprising:
- at least one ATP-releasing nucleotide (ARN) having a structure
- wherein R is where R is any purine or pyrimidine, or an analog thereof that retains an ability to base pair with a complementary nucleotide;
- and optionally dNTPs, wherein the combination of dNTPs and ARN is sufficient to provide a substrate for all bases present in the sequence of interest;
- a primer complementary to the target virus RNA;
- and
- a reverse transcriptase that incorporates ARNs;
- and
- detecting the presence of ATP released during extension of the target by the reverse transcriptase.
6 more elements
Dependencies are inferred from the claim text.