Amplification of nuceic acids using temperature zones
AssigneeKoninklijke Philips Electronics N.V.
InventorsDerk Jan Wilfred Klunder, Anke Pierik, Aleksey Kolesnychenko, Marius Iosif Boamfa, Richard Joseph Marinus Schroeders
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Abstract
The present invention relates to the amplification and detection of amplified nucleic acid sequences employing methods and devices with distinct temperature zones. The methods and devices of the present invention may be used for quantitative analysis of target nucleic acid sequences, for simultaneous quantitative analysis of multiple target nucleic acid sequences or for analyzing a sample for the presence of a target nucleic acid.
Claims26
Method for amplification and detection of target nucleic acid sequences in an amplification solution in a reaction container, comprising the steps of
- providing a reaction container comprising at least one surface hybridization zone in which capture probes are immobilized on a surface, wherein said capture probes are substantially complementary to regions on said target nucleic acid sequences;
- adding the amplification solution to said reaction container;
- generating a temperature zone profile in the reaction container with at least two kinds of thermally decoupled zones, wherein one kind of zone is identical or at least overlapping to/with the surface hybridization zones, wherein the surface hybridization zones have a generated temperature allowing for hybridization of the capture probes to the target nucleic acid sequences;
- performing an amplification of target nucleic acid sequences in the reaction container;
- and
- detecting amplified nucleic acid sequences in periodic or defined intervals during and/or after amplification, wherein amplified nucleic acid sequences are detected by hybridization of capture probes to said amplified nucleic acid sequences in said surface hybridization zone.
A device for amplification and detection of target nucleic acid sequences in an amplification solution in a reaction container, comprising
- a reaction container for receiving an amplification solution comprising said target nucleic acid sequences, wherein the reaction container comprises at least one surface hybridization zone in which capture probes are immobilized on a surface, wherein said capture probes are substantially complementary to regions on said target nucleic acid sequences and at least one other kind of zone;
- one or more temperature controllers for controlling a temperature profile of at least two kind of temperature zones in said reaction container, wherein one kind of zone is substantially overlapping with said surface hybridization zones, and wherein the surface hybridization zone has a substantially constant generated temperature allowing for hybridization of capture probes to complementary target nucleic acid sequences;
- a detection system that detects target nucleic acid sequences which are bound to said capture probes but does essentially not detect target nucleic acid sequences which are not bound to said capture probes;
- and
- a transportation system for transporting the amplification solution between the zones.
A cartridge for amplification and detection of target nucleic acid sequences in an amplification solution in a reaction container, comprising a reaction container for receiving an amplification solution comprising said target nucleic acid sequences, wherein the reaction container comprises at least one surface hybridization zone in which capture probes are immobilized on a surface, wherein said capture probes are substantially complementary to regions on said target nucleic acid sequences and wherein the reaction container further comprises at least one other kind of zone.
Dependencies are inferred from the claim text.