Patent ID: 8637302

Claim:
An apparatus comprising: a closed reactor comprising: a substrate having opposing first and second planar opposing surfaces, the substrate having a cavity and a refractive index greater than a refractive index of water; a buffer layer arranged over the first planar surface of the substrate; a cover plate arranged over the buffer layer and the cavity, the cover plate in combination with the cavity and buffer layer defining a reaction chamber; and at least one inlet port and at least one outlet port to communicate with the reaction chamber through the substrate to enable the passage of fluid from an external source into and through the reaction chamber; a temperature control system coupled to the closed reactor to cycle the temperature of a buffer solution contained within the closed reactor to enable a plurality of polymerase chain reactions, wherein the buffer solution is substantially in contact with the first surface of the substrate and being capable of sustaining a plurality of polymerase chain reactions, a plurality of hybridization reactions, and containing one or more fluorescently tagged primers, one or more optionally fluorescently tagged dNTPs, and one or more target nucleic acids; two or more target nucleic acid probes, each consisting of a short oligonucleotide with 30-50% G+C content, and with a discriminating nucleotide located near the middle of each probe; a light source coupled to the closed reactor to provide a ray of light having a wavelength chosen to activate one or more fluorescently tagged target amplicons hybridized to two or more target nucleic acid probes immobilized in independent areas on the first surface of the substrate, incident on an interface between the substrate and the buffer solution at an angle chosen to propagate an evanescent wave into the buffer solution; and a detector coupled to the closed reactor to detect the one or more fluorescent responses emitted by one of the one or more fluorescently tagged target amplicons hybridized to two or more target nucleic acid probes immobilized in independent areas on the first surface of the substrate.