Patent DE-102012203964-B3 (DE)
Title: Determining concentration of specific nucleic acids in sample, comprises providing first probe comprising first and second oligonucleotides complementary to each other, and contacting first probe and second oligonucleotide with sample
Inventor: STEHR JOACHIM
IPC: G16B25/00
Filed: 2012-03-14
Granted: 2013-02-28
Abstract: The method comprises providing a first probe comprising a first oligonucleotide (3) on its surface, providing a second oligonucleotide, which is partially complementary to the first oligonucleotide or to a first oligonucleotide adapter that is partially complementary to the first oligonucleotide and is partially complementary to a reference sequence, and contacting the first probe and the second oligonucleotide and optionally the first oligonucleotide adapter with a sample (9), where a specific nucleic acid contained in the sample is hybridized with the second oligonucleotide. The method comprises providing a first probe comprising a first oligonucleotide (3) on its surface, providing a second oligonucleotide, which is partially complementary to the first oligonucleotide or to a first oligonucleotide adapter that is partially complementary to the first oligonucleotide and is partially complementary to a reference sequence, and contacting the first probe and the second oligonucleotide and optionally the first oligonucleotide adapter with a sample (9), where a specific nucleic acid contained in the sample is hybridized with the second oligonucleotide and the second oligonucleotide is activated by hybridization. The activation of the second oligonucleotide takes place such that the first probe is bound to a second probe via the first oligonucleotide and directly via the first oligonucleotide adapter or indirectly via the second oligonucleotide or directly via a second oligonucleotide adapter. The first probe and/or the second probe contains nano-particles, where a measurable change results from one another by changing the distances of the first probe and second probes. Functional oligonucleotides having a fluorescent and/or fluorescence acceptor are modified, and contain a hairpin (8). The first probe and/or the second probe has different oligonucleotides on its surface. Independent processes are simultaneously performed in the sample, and the concentration of different types of nucleic acids are determined by a measured value that is read out at different temperatures. The first oligonucleotide adapter and/or the second oligonucleotide adapter consists of oligonucleotides that are partially complementary to each other. A further functional oligonucleotide is activated with an oligonucleotide and a nucleic acid B, by hybridization, in addition to the second oligonucleotide, where the measurable change occurs in a logical-AND-operation only when both a nucleic acid A and the nucleic acid B are present in sufficient concentration in the sample. Binary input values of the nucleic acids are logically linked in the sample so that a binary output value is created in accordance with the logic operation. The measurable change is determined before and after the excitation of the first probe and/or second probes by a light source. An independent claim is included for a kit for determining a concentration of a type of specific nucleic acids in a sample, where the sequences of the specific nucleic acids partially match to a reference sequence, and/or determining the degree of matching of the sequences of the type of specific nucleic acids with the reference sequence in a sample.