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US 10,590,472 B2Granted
Methods of detecting nucleic acids and applications thereof
OwnerZhi Zhenginventor
Life of this patent
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Abstract
The present application provides simple, specific and sensitive methods for detecting a target nucleic acid in a biological sample, detecting a pathogen in a biological sample, and diagnosing a disease in an individual, utilizing a combination of nucleic acid hybridization-based capture and ligation-enabled PCR. The methods are particularly useful for detecting low level nucleic acids and pathogens and for automation and processing of multiple biological samples.
Claims18
A method of detecting a target nucleic acid in a biological sample, comprising:
- a) capturing the target nucleic acid through a plurality of capture extenders, wherein each of the capture extenders comprises a capturing sequence that hybridizes to a region on the same target nucleic acid and an immobilizing sequence that hybridizes to a capture probe conjugated to a solid support, thereby immobilizing the target nucleic acid to the solid support;
- b) contacting the target nucleic acid with a plurality of detection probes, wherein each of the plurality of detection probes comprises a sequence that hybridizes to a region on the same strand of the same target nucleic acid;
- c) ligating the plurality of detection probes hybridized to the target nucleic acid to form a ligated detection sequence;
- d) amplifying the ligated detection sequence;
- and
- e) detecting the amplified ligated detection sequence.
3 more elements
A method of detecting a plurality of target nucleic acids in a biological sample, comprising:
- a) capturing one of the plurality of target nucleic acids through a plurality of capture extenders, wherein each of the capture extenders comprises a capturing sequence that hybridizes to a region on one of the target nucleic acids and an immobilizing sequence that hybridizes to a capture probe conjugated to a solid support, thereby immobilizing the target nucleic acid to the solid support;
- b) contacting the same one of the plurality of target nucleic acids with a plurality of detection probes comprising a first primer binding site and a second primer binding site, wherein each of the plurality of detection probes comprises a sequence that hybridizes to a region on the target nucleic acid;
- c) carrying out steps a) and b) for each of the plurality of target nucleic acids;
- d) ligating the plurality of detection probes to form a plurality of ligated detection sequences specific to each of the plurality of target nucleic acids;
- e) amplifying the plurality of ligated detection sequences;
- and
- f) detecting the plurality of amplified ligated detection sequences.
4 more elements
Dependencies are inferred from the claim text.