Immobilized crispr enriches captured target pathogens (icecap) assay chemistry key differentiators
OwnerJulie Lucas, Chelsey Smith +1inventors
InventorsJulie Lucas, Chelsey Smith, Richard Winegar
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Abstract
The present disclosure relates to an assay method for detection of target nucleic acid. The method includes immobilizing a guide RNA (gRNA) to an immobilization surface; complexing a Cas enzyme to the gRNA; adding a target nucleic acid; labeling the target nucleic acid; detecting the target nucleic acid without amplification; and determining one or more results based on the detected target nucleic acid via a reader.
Claims32
An assay method for detection of target nucleic acid, the method comprising:
- immobilizing a guide RNA (gRNA) to an immobilization surface;
- complexing a Cas enzyme to the gRNA;
- adding a target nucleic acid;
- labeling the target nucleic acid;
- detecting the target nucleic acid without amplification;
- and
- determining one or more results based on the detected target nucleic acid via a reader.
A system for detecting a target pathogen comprising:
- an immobilization surface,
- a gRNA attached to the immobilization surface,
- a Cas enzyme complexed to the gRNA, and
- a target nucleic acid derived from a pathogen.
A method of performing a multiplex assay comprising:
- immobilizing a gRNA to an immobilization surface;
- complexing a Cas enzyme to the gRNA;
- adding a target nucleic acid;
- labeling the target nucleic acid;
- detecting the target nucleic acid without amplification;
- determining one or more results based on the detected target nucleic acid via a reader;
- and wherein:
- the target nucleic acid is labeled with a Cas sandwich comprising:
- a randomized gRNA library complexed with a Cas sandwich protein;
- the randomized gRNA library comprising a plurality of secondary gRNAs;
- the secondary gRNAs comprising a tail complexed to a Cas sandwich protein and are labeled;
- and
- the randomized gRNA library comprises secondary gRNAs comprising a sequence at least 80% identical to SEQ. ID NO. 1, SEQ. ID NO. 2, or SEQ ID. NO. 3.
A method comprising:
- producing a library of gRNAs comprising a sequence at least 80% identical to SEQ. ID NO. 1, SEQ. ID NO. 2, or SEQ ID. NO. 3.
A method comprising:
- using a library of gRNAs for nucleotide detection comprising a sequence at least 80% identical to SEQ. ID NO. 1, SEQ. ID NO. 2, or SEQ ID. NO. 3.
A gRNA library comprising a sequence at least 80% identical to SEQ. ID NO. 1, SEQ. ID NO. 2, or SEQ ID. NO. 3.
Dependencies are inferred from the claim text.