Device for thermo-dependent chain reaction amplification of target nucleic acid sequences, measured in real-time
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Abstract
The present invention concerns a device for amplifying target nucleic acids, reaction cartridge s for use in the device, and modes of use of the device.
31 Citations
73 Claims
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1-51. -51. (CANCELED)
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52. A method for amplifying a nucleic acid using a device comprising at least one cartridge having a plurality of reaction chambers and a reservoir, said reaction chambers being connected to the reservoir via channels, at least one heating plate having at least two distinct zones that can be heated to at least two different temperatures and a means for relative displacement between said at least one cartridge and said at least one heating plate, allowing a cyclic variation of the temperature of the reaction chambers, said method comprising:
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a) at least partially filling said reservoir with a fluid containing a sample of nucleic acids to be analyzed and reagents for carrying out an amplification reaction, with the exception of primers, b) distributing said fluid to the reaction chambers of said at least one cartridge, in which are located the primers, c) employing said means for relative displacement between said at least one cartridge and said at least one heating plate to successively bring the contents of each reaction chamber to the at least two temperatures defined by the at least two distinct zones of said at least one heating plate. - View Dependent Claims (53, 54, 55, 56, 57, 58, 59, 60)
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61. A method for amplifying a nucleic acid using a device comprising at least one cartridge having a plurality of reaction chambers and a reservoir, said reaction chambers being connected to the reservoir via channels, at least one heating plate having at least two distinct zones that can be heated to at least two different temperatures and a means for relative displacement between said at least one cartridge and said at least one heating plate, allowing a cyclic variation of the temperature of the reaction chambers, said method comprising:
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a) at least partially filling said reservoir with a fluid containing a sample of nucleic acids to be analyzed;
b) distributing said fluid to the reaction chambers of said at least one cartridge, in which are located the primers and the reagents for carrying out an amplification reaction, c) employing said means for relative displacement between said at least one cartridge and said at least one heating plate to successively bring the contents of each reaction chamber to the at least two temperatures defined by the at least two distinct zones of said at least one heating plate. - View Dependent Claims (62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72)
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73. A process for closed system filling of reaction chambers in a cartridge comprising a plurality of reaction chambers connected to at least one reservoir via a channel having a cross section included in a circle with a diameter of less than 3 mm, the disposition of said plurality of reaction chambers and said channels with respect to said reservoir allowing a fluid to be homogeneously distributed into the plurality of reaction chambers from said reservoir, said process comprising:
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a) at least partially filling said reservoir with a fluid;
b) connecting said cartridge to means for adjusting pressure;
c) applying an underpressure inside said cartridge, then re-establishing the pressure.
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Specification