Integrated device for real time quantitative PCR

摘要

A method for real-time quantitative detection of single-type, target nucleic acid sequences amplified using a PCR in a microwell, comprising introducing in the microwell a sample comprising target nucleic acid sequences, magnetic primers, and labelling probes; performing an amplification cycle to form labelled amplicons; attracting the magnetic primers to a surface through a magnetic field to form a layer including labelled amplification products and free magnetic primers; and detecting the labelled amplification products in the layer with a surface-specific reading method.

主权项

1. A method for real-time quantitative detection of a single target nucleic acid sequence, comprising:
introducing into a microwell a sample comprising a target nucleic acid sequence, magnetic primers, and a labelled probe; performing one or more amplification cycles to form magnetic amplification products; attracting the magnetic primers to a surface through a magnetic field and forming a multilayer including the magnetic amplification products and the labelled probe hybridized thereto; and quantitatively detecting the magnetic amplification products by measuring the labelled probe in the multilayer with a spatially discriminating detection method during one or more of said amplification cycles using a spatially discriminating detection device having a vertical resolution, and the multilayer has a thickness matched to the vertical resolution of the spatially discriminating detection device.