US 6,605,436 B1Granted

Method and kit for detecting intrachromosome imbalance in interphase nuclei and their applications

AssigneesCommissariat a l'Energie Atomique, Centre National De La Recherche Scientifique, Institut Curie

InventorsKhuong Truong, Bernard Malfoy, Claire Bourgeois, Philippe Vielh, Marie-Noëlle Guilly, Bernard Dutrillaux

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Abstract

The invention concerns a method for detecting intrachromosome imbalance in interphase nuclei, characterized in that it consists in: (a) hybridizing in situ the chromosome suspected to be affected by said imbalance using two probes marked with different fluorochromes and specific of said chromosome long arm and short arm respectively; (b) measuring the intensity of the fluorescence emitted in said nuclei with different emission wavelengths corresponding to each of the two fluorochromes; (c) calculating the ratio (Ra) between the two measured values of fluorescence intensity; and (d) comparing the resulting ratio value with at least one reference value. The invention also concerns a kit for implementing said method and uses of said kit and said method, in particular for detecting cancer cells in a biological sample and for diagnosing cancer pathologies.

Claims19

Dependent claims

A method for detecting an intrachromosome imbalance in one or more interphase cell nuclei, comprising:

(a) an in situ hybridization of a chromosome present in said nuclei and suspected of being affected by said imbalance using two probes labeled with two different fluorochromes, the first probe being specific for long arms of said chromosome and the second probe is specific for short arms of the same chromosome;
(b) measuring the fluorescence intensity values emitted in said nuclei at the emission wavelengths corresponding to each of the two fluorochromes;
(c) calculating a ratio between the two fluorescence intensity values measured in step (b) (Ra);
and
(d) comparing Ra thus obtained in step (c) with at least one reference value, wherein said reference value is the ratio (Rt), wherein Rt is the ratio of two fluorescent intensity values obtained from the nuclei of control cells treated under the same conditions described in steps (a) and (b),
(e) wherein a significant difference of Ra and Rt is indicative of the intrachromosomal imbalance.

3 more elements

Dependencies are inferred from the claim text.