HIV or HCV detection with CRISPR-CAS13A
AssigneesThe J. David Gladstone Institute, a testamentary trust established under the Will of J. David Gladstone, The Regents of the University of California
InventorsParinaz Fozouni, Melanie Ott, Jennifer A. Doudna
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Abstract
The present disclosure relates to methods using CRISPR-Cas13a enzyme, complexed with HIV or HCV crRNAs to specifically and sensitively detect and quantify the presence of HIV or HCV RNA in a sample. These methods can be used to diagnose HIV or HCV infection, quantify the concentration of HIV or HCV RNA present in a sample, identify the presence of different HIV or HCV splice variants, subtypes, or mutations, and to monitor reactivation of HIV or HCV transcription.
Claims19
A method comprising:
- (a) incubating a sample containing RNA with a Cas13a protein and at least one CRISPR guide RNA (crRNA) for a period of time to form a RNA cleavage product;
- and
- (b) detecting a level of HIV or HCV RNA cleavage product with a detector,
- wherein the RNA is not reverse transcribed prior to the detecting step,
- wherein the least one crRNA is SEQ ID NOs:
- 1-8, 18-25 or 26.
A method for quantifying HIV or HCV RNA concentration comprising:
- (a) incubating a sample containing RNA with a Cas13a protein and at least one CRISPR guide RNA (crRNA) for a period of time;
- and
- (b) analyzing the sample for HIV or HCV RNA cleavage product concentration with a detector,
- wherein the RNA is not reverse transcribed prior to the analyzing step,
- wherein the least one crRNA is SEQ ID NOs:
- 1-8, 18-25 or 26.
Dependencies are inferred from the claim text.