Method and kit for diagnosing and/or quantifying by sandwich hybridisation of nucleic acid sequences on solid support

Method according to any one of the preceding claims, characterized in that a portion (13) of the target nucleotide sequence which does not hybridize with the trapping nucleotide sequence (5) and with the nucleotide sequence(s) labeled (6) or otherwise (11) is less than 60 bases, preferably less than 40 bases.

Method according to any one of the preceding claims, characterized in that the solid support (3) is a support chosen from the group consisting of tubes, filters, beads, which may be magnetic, multiwell plates or a mixture thereof.

Method according to any one of the preceding claims, characterized in that the target nucleotide sequence (2) is an amplicon resulting from a preliminary amplification by a gene amplification method, preferably chosen from the group consisting of PCR, LCR, CPR, NASBA or ICR.

Method according to any one of the preceding claims, characterized in that it also comprises a bringing into contact of the sandwich type of a standard nucleotide sequence (1) under the same conditions as the target nucleotide sequences (2) to be detected and/or quantified and comprises an additional step of quantification of the ratio between the specific labeling of the target nucleotide sequence (2) and the specific labeling of the standard nucleotide sequence (1).

Method according to any one of the preceding claims, characterized in that the sandwich hybridization is carried out in two steps.

Method according to any one of the preceding claims, characterized in that the attachment of the trapping nucleotide sequence (5) to the solid support (3) is carried out via a 5′ terminal phosphate onto an amine functional group of the solid support (3) by reaction with carbodiimide.

Method according to any one of the preceding claims, characterized in that the target nucleotide sequence (2) is a DNA.

Kit for detecting and/or quantifying, by a “sandwich”-type hybridization, a target nucleotide sequence (2) comprising a trapping nucleotide sequence (5) attached to an insoluble solid support (3) complementary to a portion (7) of the said target nucleotide sequence (2) and at least one labeled nucleotide sequence (6) complementary to another portion (8) of the target nucleotide sequence (2), characterized in that the said trapping nucleotide sequence (5) is covalently attached by one of its ends to the solid support (3) and has a length of between 50 and 300 bases, and in that the portion (10) of the trapping nucleotide sequence (5) which does not hybridize with a portion (7) complementary to the target nucleotide sequence (2) is less than 60 bases, preferably less than 40 bases, more particularly less than 20 bases.