| US 7,338,763 B2 | ||
| Method and kit for the detection and/or quantification of homologous nucleotide sequences on arrays | ||
| Jose Remacle, Malonne (Belgium); Sandrine Hamels, Ways (Belgium); Sophie Burteau, Namur (Belgium); and Nathalie Zammatteo, Gelbressee (Belgium) | ||
| Assigned to Eppendorf Array Technologies S.A., Namur (Belgium) | ||
| Filed on Jun. 02, 2004, as Appl. No. 10/860,388. | ||
| Prior Publication US 2005/0272044 A1, Dec. 08, 2005 | ||
| This patent is subject to a terminal disclaimer. | ||
| Int. Cl. C12Q 1/68 (2006.01); C07H 21/02 (2006.01); C07H 21/04 (2006.01) | ||
| U.S. Cl. 435—6 [536/23.1; 536/24.3] | 24 Claims |
| 1. A method for the identification and/or quantification of the presence of one or several among at least 7 organisms or part
thereof in a biological sample, comprising the steps of:
-amplifying a target nucleotide sequence specific for said organisms using at least 2 different primer pairs, each primer
pair being capable of amplifying nucleotide sequences from at least 4 different organisms and having a nucleotide sequence
homology higher than 85% with each of the specific nucleotide sequences to be amplified and wherein each primer of a pair
is chosen in a sequence coding for a different protein;
-providing an array onto which single-stranded capture nucleotide sequences are arranged at pre-determined locations at a
density of at least 10 different capture nucleotide sequences/cm2, said single-stranded capture nucleotide sequences being covalently bound to an insoluble support via a spacer which is at
least 20 nucleotides in length, and comprising a nucleotide sequence of about 10 to 50 bases specific for a given target sequence
of a specific organism and showing a homology of less than 85% to the other capture nucleotide sequences;
-contacting all of said target amplified sequences with the array in one solution under conditions allowing hybridization
of the target amplified sequences to complementary capture nucleotide sequences present on the array;
-detecting and quantifying signals present on specific locations on the array;
wherein the intensities of the signals at specific locations allows identification and quantification of the presence in the
biological sample of one or several among said at least 7 organisms.
|