System for charge-based detection of nucleic acids
AssigneeInfectio Recherche, Inc.
InventorsLuc Bissonnette, Frederic Raymond, Regis Peytavi
Life of this patent
Estimating the term…
Abstract
This present invention relates methods for detecting the presence of nucleic acids in a sample. In these methods, neutral capture probes are exposed to a sample possibly containing complementary nucleic acid targets. The foregoing mixture is submitted to conditions that provide for the nucleic acid targets to bind with the neutral probes thereby generating hybrids. These hybrids are submitted to positively charged reporters such as atoms, molecules or macromolecules, which electrostatically bind to the hybrids. The complexes formed between reporters and hybrids are detected by a variety of detection methods. Kits for detecting the presence of nucleic acids in a sample are also disclosed herein.
Claims58
A method for detecting the presence of nucleic acids in a sample, said method comprising:
- (a) exposing uncomplexed neutral capture probes to a sample possibly containing complementary nucleic acid targets, thereby generating a mixture;
- (b) submitting said mixture to hybridization conditions which provide for said nucleic acids targets to bind specifically to complementary neutral capture probes, thereby generating negatively charged capture probe-nucleic acid target hybrids;
- (c) submitting said negatively charged hybrids to positively charged reporters selected from group consisting of transition metal atoms, molecules, and macromolecules being capable of electrostatically binding to said hybrids, thereby generating higher-order complexes;
- and
- (d) detecting said higher-order complexes.
A method for detecting the presence of nucleic acids in a sample, said method comprising:
- (a) exposing uncomplexed neutral capture probes to a sample possibly containing complementary nucleic acid targets and containing positively charged reporters selected from group consisting of transition metal atoms, molecules and macromolecules, thereby generating a mixture;
- (b) submitting said mixture to hybridization conditions which provide for said nucleic acids targets to bind specifically to complementary neutral capture probes, thereby generating negatively charged capture probe-nucleic acid target hybrids, said reporters being capable of electrostatically binding to said hybrids, thereby generating higher-order complexes;
- and
- (c) detecting said higher-order complexes.
A method according to claim 51, wherein said enzymes comprise alkaline phosphatase having polystyrene beads conjugated thereto.
A kit for detecting the presence of nucleic acids in a sample, said kit comprising:
- uncomplexed neutral capture probes;
- a control sample possibly containing nucleic acid targets that are complementary to the neutral capture probes;
- and
- one or more positively charged reporters selected from the group consisting of transition metal cations, molecules or macromolecules;
- said reporters being capable of electrostatically binding to negatively charged capture probe-nucleic acid target hybrids.
A kit according to claim 55, wherein said support surface comprises a solid support surface 58, A kit according to claim 57, wherein said solid support surface comprises a probe array.
Dependencies are inferred from the claim text.