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US 6,790,623 B2Granted
DNA detection by means of a strand reassociation complex
AssigneeRoche Diagnostics GmbH
InventorsKurt Weindel, Joachim Brand
Life of this patent
Estimating the term…
Abstract
The discriminating capability of hybridization assays is increased by a combination of labelled primers which produce amplificates of one strand of a nucleic acid with a capture probe which is complementary to the same strand of the nucleic acid.
Claims11
A method for the detection of a nucleic acid comprising the steps:
- a) amplifying the nucleic acid or a part thereof in a reaction mixture with a first primer and a second primer, one of which can hybridize with one strand of the nucleic acid to be detected and the other of which can hybridize with a strand complementary thereto, wherein one of the primers contains a detectable label, to form a labeled extension product and an unlabeled extension product which are complementary to each other;
- b) contacting a capture probe with the extension products to form a hybridization complex wherein the complex is necessarily a duplex, said hybridization complex comprising the capture probe, the labeled extension product and the unlabeled extension product, wherein the capture probe is selected such that the capture probe hybridizes to only the unlabeled extension product and only the unlabeled extension product hybridizes with the labeled extension product;
- c) separating the hybridization complex from other components of the reaction mixture;
- and
- d) determining the detectable label in the hybridization complex of step b).
2 more elements
A method for the selective detection of specific sequences in one or several nucleic acids(s) from a sample comprising the steps:
- a) amplifying the nucleic acids or parts thereof in a reaction mixture with at least one set of two primers, wherein a first primer in each set of primers can hybridize with one strand of the nucleic acid(s) and a second primer in each set can hybridize with a strand complementary thereto, wherein one of the first and second primers contains a detectable label, to form a labeled extension product and an unlabeled extension product which are complementary to each other for each of the specific sequences;
- b) contacting at least one capture probe with the extension products to form hybridization complexes wherein the complexes are necessarily duplexes, said hybridization complexes comprising a capture probe, the labeled extension product and the unlabeled extension product for each of the specific sequences, wherein the capture probe is selected such that the capture probe hybridizes to only the unlabeled extension product and only the unlabeled extension product hybridizes with the labeled extension product;
- c) separating the hybridization complexes from other components of the reaction mixture;
- and
- d) determining the detectable label in the hybridization complexes of step b).
2 more elements
Dependencies are inferred from the claim text.