Real time-PCR of targets on a micro-array

摘要

The present invention relates to a method and apparatus for monitoring on a micro-array a PCR amplification of a nucleotide molecule being present in a solution. The method comprises the steps of: providing a support (15) having fixed upon its surface a microarray comprising at least a capture molecule (20) being immobilized in specifically localized areas (21) of said support and a reaction chamber (14) on; introducing a solution containing said nucleotide molecule into said reaction chamber (14) and reagents for nucleotide molecule amplification and labelling; submitted the solution to at least 2 thermal cycles having at least 2 and preferably 3 different temperature steps in order to obtain labelled target nucleotide molecule (13) by PCR amplification; performing at least a measurement of the labelled target nucleotide molecule in at least one thermal cycle by incubating said labelled target nucleotide molecule (13) under conditions allowing a specific binding between said target nucleotide molecule (13) and its corresponding capture molecule (20) and measuring the light emission (7) from the bound labelled target nucleotide molecule in response to excitation light (2) with said solution being present in the chamber and containing the labelled target nucleotide molecule (13), wherein the surface of emission for a localized area is comprised between about 0.1 µm 2 and about 75 mm 2 , and; Processing the data obtained in at least one thermal cycle in order to detect and/or quantify the amount of nucleotide molecule present in the solution before the amplification.