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OwnerJose Remacle, Muriel Art +2inventors
InventorsJose Remacle, Muriel Art, Laurence Lockman, Nathalie Zammatteo
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The present invention relates to a method for the identification and/or the quantification of one or more target nucleotide sequences (3) present homologous to each other and being possibly present in a sample and discrimination from possibly homologous sequences, by using two sets of nucleotide sequences (1 and 2), wherein a first set of nucleotide sequences possibly labelled (1) bind specifically to said target nucleotide sequences (3) in a first step and are detected and/or quantified through hybridisation with a second set of capture nucleotide sequences (2) having at least a part of sequence complementary to the possibly labelled nucleotide sequence (1) of the first set of nucleotide sequences, said capture nucleotide sequence (2) being immobilised upon the surface (4) of a solid support according to an array of at least 4 discrete regions/cm2, each of said discrete regions being bonded with one species of capture nucleotide sequences (2), and wherein the identification and quantification of the binding between the possibly labelled nucleotide sequence (1) and their corresponding capture nucleotide sequence (2) is correlated with the identification and the quantification of a target nucleotide sequence (3) present in the sample.
Method for the identification and/or the quantification of one or more target nucleotide sequences (3) present homologous to each other and being possibly present in a sample and discrimination from possibly homologous sequences, by using two sets of nucleotide sequences (1 and 2), wherein a first set of nucleotide sequences possibly labelled (1) bind specifically to said target nucleotide sequences (3) in a first step and are detected and/or quantified through hybridisation with a
The method according to any of the preceding claims, wherein the target nucleotide sequences (3) to be identified and/or quantified present an homology with homologous sequences present in the sample higher than 30%, preferably higher than 60%, more preferably higher than 80%.
The method according to any of the preceding claims, wherein the target nucleotide sequences (3) to be identified and/or quantified differ from homologous sequences by only one base.
The method according to any of the preceding claims, wherein the possibly labelled nucleotide sequences (1) contain a part of a portion of their sequences comprised between 10 and 60 bases, a sequence which is specific to the target nucleotide sequence (3).
The method according to any of the preceding claims, wherein the capture nucleotide sequences of the second set of nucleotide sequences present a sequence which is complementary to the possibly labelled nucleotide sequences (1).
The method according to any of the preceding claims, wherein the possibly labelled nucleotide sequences (1) contain part of their sequence being not specific of the target but specific of their capture probes.
The method according to any of the preceding claims, wherein the labelled nucleotide sequences (1) contain a mixture of two or more labelled sequences terminated by a different nucleotide which binds to the target sequence adjacent to the possible SNP, one of the end nucleotide being perfectly matched to the SNP, and another annexed probe which will bind to the other site of the SNP target, the perfectly matched probe being ligated to this probe by a ligase before detected on the capture probe array.
The method according to any of the preceding claims, wherein the labelled nucleotide sequences (1) contain a mixture of probes each one differing in at least one base located at the SNP site and treated with a specific nuclease before being detected on the capture probe array.
The method according to any of the preceding claims, wherein the one or more target nucleotide sequences (3) to be detected and/or quantified are rRNAs, preferably selected from the group consisting of 16S, 23S, 18S and 25S rRNAs.
The method according to any of the preceding claims, wherein the solid support is selected from the group consisting of glass, electronic device, silicon support, plastic support, compact disc, filter, metallic support, polylysine coated surfaces or a mixture thereof.
A diagnostic and/or quantification apparatus of one or more homologous target nucleotide sequences possibly present in a sample, which comprises means to perform the method according to any of the preceding claims.
Dependencies are inferred from the claim text.