Device and method for multiplexed detection of nucleic acid sequences

Abstract

A device for multiplexed detection of nucleic acid sequences comprises a thermal cycler arranged to perform a series of thermal cycles with an in vitro nucleic acid extraction reagent containing at least two fluorescence probes combining a quencher and a fluorophore. The fluorescence probes are each arranged to target a distinct nucleic acid sequence and the fluorophores have overlapping fluorescence wavelength ranges. A light sensor is arranged to measure radiation emitted by the fluorophores in the fluorescence wavelength ranges, the radiation emitted by each probe varying depending on whether the latter is in a non-modified state in which the quencher mitigates, or does not substantially mitigate, the fluorescence emission, or in a modified state in which the quencher has an opposite effect on the fluorescence emission. An analyzer is arranged to determine, for each respective fluorescence probe, a value representative of a concentration in a modified state from time signatures derived from the measured fluorescence as a function of time for a given thermal cycle of a reaction mixture comprising one or more probes, which may each be substantially entirely in a non-modified or modified state, and at least two measurements carried out during at least some of the thermal cycles, which values representative of a concentration in a modified state make it possible to qualify the presence of one or more nucleic acid sequences each associated with a distinct fluorescence probe so as to cause the fluorescence probe to change from the non-modified state to the modified state during interaction.

Dependencies are inferred from the claim text.