Abstract: Provided herein, in certain embodiments, are various methods, reagents, and devices for detection of multiple target nucleic acids in a sample, or multiple segments of a target nucleic acid in a sample, using a programmable nuclease. In certain embodiments, the present disclosure provides compositions of pools of guide nucleic acids, programmable nucleases, and detector nucleic acids and methods of using said compositions for detection of different segments of one target nucleic acid or different target nucleic acids in a sample.

Abstract: Described herein, in certain embodiments, are methods of assaying for a segment of a coronavirus target nucleic acid in a sample. In some embodiments, the target nucleic acid is from a gene of SARS-CoV-2, or a variant thereof. In some embodiments, methods described herein comprise a) contacting the sample to: i) a detector nucleic acid; and ii) a composition comprising a programmable nuclease and a non-naturally occurring guide nucleic acid, and b) assaying for a change in a signal, wherein the change in the signal is produced by cleavage of the detector nucleic acid by the programmable nuclease. Also provided herein are devices for performing the methods of assaying for a segment of a coronavirus target nucleic acid described herein.