Chip-based isothermal amplification devices and methods
AssigneeThe Regents of the University of Michigan
InventorsMark A. Burns, David T. Burke, Brian N. Johnson, John D. DeNuzzio, Wayne F. Beyer JR.
Life of this patent
Estimating the term…
Abstract
Disclosed are methods and compositions for isothermal amplification of nucleic acids in a microfabricated substrate. Methods and compositions for the analysis of isothermally amplified nucleic acids in a microfabricated substrate are disclosed as well. The microfabricated substrates and isothermal amplification and detection methods provided are envisioned for use in various diagnostic methods, particularly those connected with diseases characterized by altered gene sequences or gene expression.
Claims62
An apparatus for use in the isothermal amplification of a selected nucleic acid, comprising a microfabricated substrate defining at least a first channel, said at least a first channel connected to a reaction chamber, and a means for isothermally regulating the temperature of said reaction chamber.
A method for isothermal amplification of a selected nucleic acid, comprising:
- a) providing a sample comprising said selected nucleic acid, and reagents effective to permit an isothermal amplification reaction, to a microfabricated substrate that defines at least a first channel, said at least a first channel connected to an isothermally regulated reaction chamber;
- and
- b) conducting an isothermal amplification reaction to amplify said selected nucleic acid.
A method for detecting the presence of a selected nucleic acid, comprising:
- a) introducing a sample suspected of containing said selected nucleic acid, and reagents effective to permit an isothermal amplification reaction, into a microfabricated substrate defining at least a first channel, said at least a first channel connected to an isothermally regulated reaction chamber;
- b) conducting an isothermally regulated amplification reaction to amplify said selected nucleic acid;
- and
- c) detecting the presence of the amplified selected nucleic acid, wherein the presence of the amplified selected nucleic acid confirms the presence of said selected nucleic acid in said sample.
A method of making a device for use in isothermal nucleic acid amplification, comprising preparing at least a first microfabricated device, chip or wafer defining at least a first channel that is operably connected to an isothermally regulated reaction chamber.
A kit for conducting isothermal amplification of a selected nucleic acid, comprising, in suitable container means:
- a) at least a first microfabricated substrate defining at least a first channel, said at least a first channel connected to an isothermally regulated reaction chamber;
- and
- b) reagents effective to permit an isothermal nucleic acid amplification reaction.
A diagnostic system for identifying a selected nucleic acid, comprising at least a first microfabricated substrate defining at least a first channel that is connected to at least a first isothermally regulated reaction chamber;
- said diagnostic system further comprising a nucleic acid analysis component and a nucleic acid detection means in operable association with the reaction chamber of said at least a first microfabricated substrate.
A method of making a nucleic acid diagnostic system, comprising preparing at least a first microfabricated substrate defining, in a series of operable associations, at least a first channel, an isothermally regulated reaction chamber, a nucleic acid analysis component and a nucleic acid analysis detection means.
Dependencies are inferred from the claim text.