Eppendorf Array Technologies S.a.

Eppendorf Array Technologies

Sylvain Margaine

Dieter Husar

Isabelle Alexandre

José Remacle

Heinz Koehn

Nathalie Zammatteo

The present invention relates to a method and apparatus for monitoring a real-time quantification of multiple target molecules during their binding on capture molecules of a micro-array. The method comprises the steps of: placing, in a chamber (), a support () having fixed upon its surface a micro-array comprising at least 5 capture molecules () being immobilized in specifically localized areas () of said support; introducing said labeled target molecules solution () into the chamber; incubating said labeled target molecules under stable and controlled temperature conditions to allow the binding between said target and capture molecules; directing an excitation light () from a light source () on the surface of the micro-array; measuring the electromagnetic light emission () from the bound target molecules in response to said excitation light in presence of the solution containing the target molecules wherein the surface of emission for a localized area is comprised between about 0.1 &#x3bc;mand about 10 mmand wherein each of the at least 4 localized areas is monitored with time with at least two measurements being done per localized area (), and processing and storing the values of the different measurements and quantifying at least 4 different target molecules present in solution using at least one measurement value for each said target.

Real-time quantification of multiple targets on a micro-array

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 includes the steps of: providing a support having fixed upon its surface a microarray having at least a capture molecule being immobilized in specifically localized areas of the support and a reaction chamber; introducing a solution containing the nucleotide molecule into the reaction chamber and reagents for nucleotide molecule amplification and labelling; submitting 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 by PCR amplification; performing at least a measurement of the labelled target nucleotide molecule in at least one thermal cycle by incubating the labelled target nucleotide molecule under conditions allowing a specific binding between the target nucleotide molecule and its corresponding capture molecule and measuring the light emission from the bound labelled target nucleotide molecule in response to excitation light with the solution being present in the chamber and containing the labelled target nucleotide molecule. The surface of emission for a localized area is between about 0.1 &#x3bc;mand about 75 mm. The method further includes 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.

Real-time PCR of targets on a micro-array

Disclosed is a system and method conducting real-time PCR. Unlabeled capture molecules of a specific design are immobilized on a solid support, and contacted with amplicons produced in one or more PCR cycles. Detection of amplicons may take place during or between the PCR cycles while the solid support is in fluidic contact with the PCR solution. In an alternate embodiment detection of the amplicons takes place when the solid support is not in fluidic contact with the PCR solution. The method is suitable for the simultaneous detection and quantification of closely homologous target molecules.

Identification and quantification of a plurality of biological (micro)organisms or their components

Disclosed is a system and method conducting real-time PCR. Capture molecules of a specific design are immobilized on a solid support, and contacted with amplicons produced in one or more PCR cycles. Detection of amplicons may take place during or between the PCR cycles while the solid support is in fluidic contact with the PCR solution. In an alternate embodiment detection of the amplicons takes place when the solid support is not in fluidic contact with the PCR solution. The method is suitable for the simultaneous detection and quantification of closely homologous target molecules.

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Namur, Belgium

Sylvain Margaine