METHODS AND SYSTEMS FOR ANALYSING HYBRIDISATION

摘要

A method is described for the analysis of hybridisation between a target in solution and a probe bound at a surface. The method comprises receiving detection intensity results for hybridisation of the target with a plurality of different probes, the probes being selected so that a range of hybridisation detection intensity results for the hybridisation between the target and the probe is covered. The method further comprises analysing the detection intensity results as function of the hybridisation free energy. According to embodiments of the present invention, the receiving and/or analysing takes into consideration a thermodynamic non-equilibrium state for the target-probe bounding state.

主权项

1. A method for determining in a sample solution the presence or concentration of a target nucleic acid out of a set of possible target nucleic acids, wherein each of said possible target nucleic acids differ from each other by one or two nucleotides, the method comprising
contacting said sample solution with a plurality of different probes bound at a surface and receiving detection intensity results for hybridisation of the possible target nucleic acids and each of said plurality of different probes, wherein for each possible target nucleic acid, at least one probe comprises a strand of nucleic acid that is perfectly complementary with said possible target nucleic acid and at least one other probe comprises a strand of nucleic acid that is non-perfectly complementary with said possible target nucleic acid so that a range of hybridisation detection intensity results for the hybridisation between each possible target nucleic acid and the probe is covered, for each possible target nucleic acid performing the analysis of the detection intensity results in a processor of the computing system as a function of hybridisation free energy, wherein the analysis comprises:
determining the hybridisation free energy for the hybridisation between a possible target nucleic acid in solution and a probe bound at a surface based on a three state model wherein a bound state, an unbound state and a thermodynamic non-equilibrium state for the hybridisation are taken into account;analysing the logarithm of the detection intensity results as a function of the difference between the hybridisation free energy of said hybridization with respect to the hybridisation free energy for the perfectly complementary probe of the possible target nucleic acid; andderiving the presence and/or concentration of a target nucleic acid in said sample by determining whether said relationship between the logarithm of the detection intensity results and the hybridisation free energy corresponds to a predetermined relationship between the logarithm of the detection intensity results and the hybridisation free energy between the target and said probe.