Methods for detecting multiple nucleic acids in a sample using reporter compounds and binding members thereof

A method of determining the presence and/or amount of nucleic acids in a sample, comprising

• providing in a reaction chamber an amount of a first nucleic acid;
• an amount of a second nucleic acid;
• an amount of a FRET compound capable of forming complexes with the first nucleic acid, wherein the FRET compound comprises a binding portion specific to a region of the first nucleic acid, and a first detectable label that has, in an uncomplexed state, a first detectable characteristic and, in a complexed state, a second detectable characteristic;
• and an amount of a reporter compound capable of forming complexes with the second nucleic acid, wherein each reporter compound comprises a binding portion specific to a region of the second nucleic acid and a second detectable label, wherein said second detectable label is identical to said first detectable label;
• wherein the reaction chamber comprises a binding
• member capable of capturing the reporter compound, wherein forming of complexes of the second nucleic acid with the reporter compound inhibits capturing of the reporter compound by the binding member, and a cover element which is configured to be at least partially deformable such that the reaction chamber has a relaxed state interior volume and a compressed state interior volume;
• subjecting the first and/or second nucleic acid to an isothermal amplification;
• forming complexes each comprising a first nucleic acid and a FRET compound;
• forming complexes of a subset of the amount of reporter compound with a second nucleic acid;
• capturing a remaining subset of the amount of reporter compound not in complex with a second nucleic acid on the binding member;
• determining a value indicative for the presence and/or amount of first nucleic acid in complex with FRET compounds in solution in the reaction chamber having a relaxed state interior volume;
• and
• determining a value indicative for the presence and/or amount of reporter compound captured on the binding member in the reaction chamber having a compressed state interior volume, and, based on the value indicative for the presence and/or amount of reporter compound captured on the binding member, determining a value indicative for the presence and/or amount of second nucleic acid.

A method of determining the presence and/or amount of nucleic acids in a sample, comprising

• introducing into a reaction chamber an amount of a first nucleic acid;
• an amount of a second nucleic acid;
• an amount of a FRET compound capable of forming complexes with the first nucleic acid, wherein the FRET compound comprises a binding portion specific to a region of the first nucleic acid, and a first detectable label that has, in an uncomplexed state, a first detectable characteristic and, in a complexed state, a second detectable characteristic;
• and an amount of a reporter compound capable of forming complexes with the second nucleic acid, wherein each reporter compound comprises a binding portion specific to a region of the second nucleic acid and a second detectable label, wherein said second detectable label is identical to said first detectable label;
• wherein the reaction chamber comprises a first
• surface, a second surface and a binding member capable of capturing the reporter compound, wherein the distance between the first and the second surface can be varied such that the reaction chamber has a relaxed state interior volume and a compressed state interior volume and wherein forming of complexes of the second nucleic acid with the reporter compound inhibits capturing of the reporter compound by the binding member;
• subjecting the first and/or second nucleic acid to an isothermal amplification;
• forming complexes each comprising a first nucleic acid and a FRET compound;
• forming complexes of a subset of the amount of reporter compound with a second nucleic acid;
• capturing a remaining subset of the amount of reporter compound not in complex with a second nucleic acid on the binding member;
• determining a value indicative for the presence and/or amount of first nucleic acid captured by FRET compounds in solution in the reaction chamber having a relaxed state interior volume;
• and
• determining a value indicative for the presence and/or amount of reporter compound captured on the binding member in the reaction chamber having a compressed state interior volume, and, based on the value indicative for the presence and/or amount of reporter compound captured on the binding member, determining a value indicative for the presence and/or amount of second nucleic acid.