Detection method of specific sequences of nucleic acids (versions), and device for its implementation

FIELD: biotechnologies.

SUBSTANCE: as per the first version, a method is implemented by recording of cyclic voltamperograms of a working electrode modified with carbon nanotubes with a oligonucleotide probe noncovalently immobilised on their surface, before and after the nucleic acid specimen is added to the investigated solution, and as per the change of capacitive characteristic, it is evaluated whether a section complementary to the oligonucleotide probe is available in the specimen or not. The second version of the method differs by the fact that non-covalent immobilisation of the oligonucleotide probe onto the surface of nanotubes is performed by means of an anchor group pre-introduced to the probe. This version includes recording not only of the change of surface area of voltamperograms from cycle to cycle, but also occurrence of a specific peak on a cyclic voltamperogram, which is related to fixation of detected nucleic acid complete with the modified probe. Intensity of the peak on the cyclic voltamperogram is pro rata to concentration of the determined nucleic acid, which allows performing quantitative evaluation. A device for implementation of the detection method of specific sequences of nucleic acids represents an electrochemical analyser that consists of a three-electrode electrochemical cell, the electrodes of which are connected to a recording device, and the working electrode is made from a silicone substrate modified with vertically oriented carbon nanotubes with an immobilised oligonucleotide probe complementary to the nucleic acid to be determined.

EFFECT: improving evaluation accuracy.

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The invention relates to the field of molecular biology and electrochemistry and can be used for detection of specific nucleic acids sequences (NK) (DNA or RNA) by electrochemical method using modified electrodes, in particular modified carbon nanomaterials.

In recent years, much attention is paid to the development of diagnostic PC-sensors based on carbon nanotubes (CNTS) [Wang, J,. Nanomaterial-based electrochemical biosensors // Analyst. 2005 V.130. P.421-426. Mercosi, A., Pumera, M., Llopis, X., Perez, C., Valle, M., Alegret, S. New materials for electrochemical sensing. // Trends Anal. Chem. 2005. V.24. P.836-838.], designed for the detection of infections and genetic disorders that cause the development of human diseases [Balasubramanian, K., Burghard, M. Biosensors based on carbon nariotubes // Anal. Bioanal. Chem. 2006. V.385. P.452-468; Pumera, M., Sanches, S., Ichinose, I., Tang, J. Electrochemical nanobiosensors // Sens. Actiators. 2007. V.123. P.1195-1205]. The recognition system of the biosensor based on the specific interaction of biomolecules or biological supramolecular structures. To convert the process of recognition of biomolecules in the analytical signal element of biological recognition must be in direct spatial contact with a transducer [K. Balasubramanian, M. Burghart Biosensors based on carbon nanotubes // Anal. Bioanal Chem. 2006, V.385. P.452-468], thus generated by the biosensor signal functionally related to the amount of ODA is the shared component. Highly organized structure and a variety of electronic properties of carbon nanotubes make promising use of this material for creating such biosensors as both structural and transformative elements [Mercosi, A., Pumera, M., Llopis. X:; Perez, C., Valle, M., Alegret, S. New materials for electrochemical sensing. // Trends Anal. Chem. 2005. V.24. R-838].

A promising direction in the creation of biosensors is the development of electrochemical biosensors with high selectivity and sensitivity [Wang, J., Survey and Summary. From DNA biosensors to gene chip. // Nucleic Acids Res. 2000. V.28. P.3011-3016]. Known methods of electrochemical detection of biomolecules on a change in resistance of the electrode in the specific binding of biomolecules NC-probes (oligonucleotide probes)immobilized on the surface of carbon nanotubes (CNTS), which are included in the conductive composition system [Carbon nanotube biosensors with aptamers as molecular recognition elements and method for sensing target material using the same. H.M. So, J.O. Lee, Y.H. Kim, K.H. Won, H.J. Chang, K.J. Kong, Y.M. Choi. US2008/0094078 A1. 2008. (US 7854826 B2, 2010); Sensors employing single-walled carbon nanotubes. M.S. Strano, S. Baik, P.Barone. US 2007/0292896 Al. 2007]. Describes the use of the composites of the array of oriented carbon nanotubes (CNTS), chemically attached to the substrate, made of aluminum,gold or silicon oxide, with covalently attached oligonucleotide probes for electrochemical detection PC [Berti F., Lozzi ., I. Palchetti, S. Santucci, G. Marrazza Electrochim. Acta. 2009, V.54. P.5035-5041; Arumugam P.U., Chen H., Siddiqui, S., Weinrich J.A.P., Jejelowo A., Li J., Meyyappan M.. Biosens. Bioelectron. 2009, V.24. P.2818-2824; Pumera, M., Sanchez, S., I. Ichinose, J. Tang Sens. Actuat. B. 2007, V.123. P.1195-1205]. As a signal of hybridization oligonucleotide probe and detectable nucleic acid (NC) use the values of current change.

Known method of detecting DNA prototype of the invention [S.G. Wang, R. Wang, P.J. Sellin, Zhang Q. DNA biosensors based on self-assembled carbon nanotubes // Biochem. Biophys/. Res. Commun. 2004. V.325, No. 4. P.1433-1437.], where for the detection of 21-tier DNA fragment using methods cyclic voltammetry and differential pulse voltammetry electrochemical analyzer representing a three-electrode electrochemical icaco, the electrodes of which are connected to a recording device. CNT-containing working electrode of the electrochemical analyzer produced by growing vertically oriented multiwall carbon nanotubes by catalytic deposition from the gas phase on the electrode representing a gold foil substrate coated with a layer of catalytic particles of Nickel. CNT-containing electrode is treated with concentrated nitric acid to remove catalyst particles of Nickel and oxidation of the nanotube ends. It formed during the oxidation at the ends of the nanotubes carboxyl groups covalently p is soedinyaet 21-tier

oligodeoxyribonucleotide probe containing at the 5'end of the linker with the primary amino group. Measurement of electrochemical properties hold, giving a linear scan of the potential on the electrodes. Detection is performed by comparing the peaks obtained on voltamperometry in the area from 0 to - 0.5 C.

The disadvantages of this method and device are the high cost and complexity of the receiving electrodes. Such modified electrodes have several disadvantages: low stability, the technical complexity of the introduction of the oligonucleotide probe to the surface of the CNTS, low content of probe molecules in the structure of the electrode, and, as a consequence, a relatively low sensitivity, high cost of gold foil background.

The objective of the invention is to simplify, reduce costs and accelerate detection of specific sequences NK (DNA or RNA).

The problem is solved in two variants of the method and the device.

In the first scenario the problem is solved in that in the method of detection of specific sequences of nucleic acids by electrochemical method, the detection is carried out by registration cyclic voltamperometry working electrode modified with carbon nanotubes with ecovalence immobilized on their surface oligonucleotide probe, before andafter making in the analyzed sample solution of nucleic acid (RNA or DNA), to change the capacitive characteristics defined by the area of cyclic voltamperometry, make a conclusion about the presence or absence in the sample of nucleic acid (RNA or DNA) of the site, a complementary oligonucleotide probe, after registration of cyclic voltamperometry working electrode before and after inclusion in the study sample solution of nucleic acid (RNA or DNA) to build a profile of changes in the relative capacity of the electrode from the rooms of the loop.

According to the second variant of the method, the task is solved in that in the method of detection of specific sequences of nucleic acids by electrochemical method, the detection is carried out by registration cyclic voltamperometry working electrode modified with carbon nanotubes with ecovalence immobilized on the surface of carbon nanotubes oligonucleotide probe containing the remainder of polyaromatic hydrocarbons as anchor groups for immobilization of the probe, before and after inclusion in the study sample solution of nucleic acid (RNA or DNA), the change in the capacitive characteristics defined by the area of cyclic voltamperometry, or by the presence or absence of a peak capacitive characteristic, make a conclusion about the presence or absence in the sample of nucleic acid (RNA is whether DNA) plot complementary oligonucleotide probe, and determine the concentration of detectable NC, after registration of cyclic voltamperometry working electrode before and after inclusion in the study sample solution of nucleic acid (RNA or DNA) to build a profile of changes in the relative capacity of the electrode from the rooms of the cycle, and oligonucleotide probe modify the remnants of pyrene or other polyaromatic hydrocarbons.

The problem is solved in that the device for implementing the method of detection of specific sequences of nucleic acids by electrochemical method is an electrochemical analyzer, consisting of a three-electrode electrochemical cell, the electrodes of which are connected to a recording device containing a power source, for example, a potentiostat, and a working electrode made of conductive silicon or metal substrate with grown therein vertically oriented carbon nanotubes, on the entire surface of which ecovalence immobilized oligonucleotide probe, complementary to the area defined NC, or modified oligonucleotide probe, complementary to the area defined NC, between the working and auxiliary electrode is separator, with the oligonucleotide probe prewar the positive modify polyaromatic hydrocarbon, which acts as an anchor group for non-covalent immobilization of the modified oligonucleotide probe on the surface of the carbon nanotubes of the working electrode, as well as oligonucleotide probe modify the remnants of pyrene or other polyaromatic hydrocarbons.

Figure 1 presents the device to implement the method. All elements of the three-electrode cells are located on the base (1), for example PTFE, on the basis of the current collector is placed auxiliary electrode (2), made for example of titanium with iridium coating, it summed up the reference electrode (3J, for example, silver chloride, and the working electrode (4), nanotubes at the working electrode is oriented perpendicular to the plane of the silicon substrate of the working electrode. The reference electrode and the working electrode concern separator (5), for example, from non-woven polypropylene fibers, pokryvalah auxiliary electrode. The working electrode and the reference electrode are detected using a tripod (6). The current collectors of all electrodes are connected to a recording device, for example, a potentiostat Elins P-30S (7).

Pre-receive CNT-modified working electrode, representing elektroprovodyashchie silicon or a metal substrate with grown therein vertically oriented of magoten the mi carbon nanotubes. During the next 18-24 h conduct non-covalent, unlike the prototype, the immobilization specially synthesized oligonucleotide probe, complementary to the plot detectable NC, by incubation of the CNT-modified electrode in 200 μl of an aqueous solution of probe concentration 1-5·10^-5M as an oligonucleotide probe, unlike the prototype, using not only oligodeoxyribonucleotide, but oligoribonucleotide and their synthetic modified analogues (oligo(2'-O-methylribonucleotide), LNA-modified oligonucleotides and others), as well as oligonucleotide probes containing the remainder of polyaromatic hydrocarbons. Modified electrodes are dried and used in the composition of the three-electrode electrochemical cell for the specific detection of NK.

To measure the electrochemical characteristics of the electrode by the method of cyclic voltammetry with linear scan of potential in three-electrode cell on the basis of (1) put the current collector auxiliary electrode (2), an auxiliary electrode placed two layers of separator (5) from non-woven polypropylene fibers. The separator excessively impregnated with the electrolyte solution, the electrode (3) and working electrode (4) with an array of carbon nanotubes concern separator (5). The electrodes connect to registriruutsya device (7).

The method of measurement is to feed on CNT-modified working electrode with immobilized thereon oligonucleotide probe voltage varying according to a given law (linear scan of the potential), and recording the current in the circuit. Cyclic voltamperometry in the potential window for measurements [-0.2; 1] register using a potentiostat Elms R-30S, working in potentiodynamic mode. The potential is measured relative to silver chloride reference electrode. The scan speed capacity is 20 mV/surabachi electrode cycleroute at room temperature in the electrolyte of the following composition: 0.1 M NaCl, 10 mm cacodylate sodium, 1 mm Yahata, pH 7.4. After 3 cycles in the electrochemical cell, add a solution of the NC in the same buffer and continue Cycling. As the study NC used long synthetic RNA fragments. After the introduction of the sample see the change in cyclic voltamperometry, namely, the change of the area of voltamperes from cycle to cycle. Form received voltamperes corresponds to the electrochemical processes in the cell, and the average current is proportional to the surface area of the electrode material. The findings build cyclic voltamperometry and calculate the pixel electrode in each of the loop. When the increase/reduction of capacity during Cycling is possible to draw conclusions about the status and activity of the electrode surface. On the basis of these data, draw conclusions about the presence or absence in the sample NC area, complementary to the oligonucleotide probe. In addition, the change of the area of voltamperes possible to estimate the concentration of NC.

When non-covalent immobilization of oligonucleotide probe on the surface of the vertically oriented CNTS via anchor groups (the second option) register not only the change of the area of voltamperes from cycle to cycle, but also the emergence of a specific peak in the cyclic voltamperometry associated with fixation detectable NK in complex with modified probe on the surface of the CNT electrode (figure 2). The intensity of the peak of the cycle voltamperometry proportional to the concentration determined NC, which allows for a quantitative assessment.

Distinctive features of the invention in the first embodiment are:

- use as probes of oligodeoxyribonucleotides or oligoribonucleotides, and their modified analogues (oligo(2'-O-methylribonucleotide), LNA and others);

- registration of cyclic voltamperometry working electrode before and after inclusion in the study sample solution NC (RNA or DNA), the soda the containing section, complementary oligonucleotide probe, ecovalence immobilized on the surface of the carbon nanotubes of the working electrode;

- check the presence or absence in the sample of NC (RNA or DNA) of the site, complementary to the oligonucleotide probe, the change in the capacitive characteristics defined by the area of cyclic voltamperometry.

Distinctive features of the invention according to the second variant are:

- use as probes of oligodeoxyribonucleotides or oligoribonucleotides, and their modified analogues (oligo(2'-O-methylribonucleotide), LNA and others);

- preliminary modification of the oligonucleotide probe polyaromatic hydrocarbon as an anchor group for non-covalent immobilization of oligonucleotide probe on the surface of the carbon nanotubes of the working electrode;

- modification of the oligonucleotide probe pyrene residues or other polyaromatic hydrocarbons;

- registration of cyclic voltamperometry working electrode modified with carbon nanotubes with ecovalence immobilized on the surface of carbon nanotubes oligonucleotide probe containing the remainder of polyaromatic hydrocarbons as the anchor group, before and after inclusion in the study sample solution NC (RNA or the NC), containing section, complementary to the probe;

- check the presence or absence in the sample of NC (RNA or DNA) of the site, complementary to the oligonucleotide probe, the change in the capacitive characteristics defined by the area of cyclic voltamperometry, or by the presence or absence of a peak voltamperometric.

- determination of the concentration of detectable NK intensity peak of the cycle voltamperometry.

Distinctive features of the device:

- working electrode is a conductive silicon or a metal substrate, a modified vertically oriented carbon nanotubes;

all over the surface of carbon nanotubes ecovalence immobilized molecules oligonucleotide probe, complementary to the defined area detectable NK;

- oligonucleotide probe pre-modified aromatic hydrocarbon, which acts as an anchor group for immobilization of oligonucleotide probe on the surface of the carbon nanotubes of the working electrode;

- oligonucleotide probe modified residues pyrene or other polyaromatic hydrocarbons;

- between the working and auxiliary electrode is a separator.

A typical example of No. 1

1. Receiving modified the working electrode

Arrays oriented multiwall carbon nanotubes grown well-known aerosol CVD method on silicon substrates with a size of 5×5 mm by decomposition of a 2%aqueous solution of ferrocene in toluene at a temperature of 800°C [A.G. Kudashov, Kuren A.G., Okotrub AV, Guselnikov, A.V., D. B.C., Bulusheva G. Synthesis and structure of films of carbon nanotubes oriented perpendicular to the substrate // ZH. 2007. T, No. 12. Pp.96-100. Okotrub AV, Bulusheva L.G., A.G. Kudashov, Belavin CENTURIES, Komogortsev S.V. Arrays of carbon nanotubes oriented perpendicular to the substrate: the anisotropy of the structure and properties. // Nanotechnologies in Russia. 2008. V.3. Issue 3-4. S-131.]. One side of the silicon substrate and the side surface of the substrate is thoroughly cleaned from carbon material, to clean silicon surface is attached to the contact conductive glue.

2. Synthesis of oligonucleotide probe of earth I

Obtaining oligonucleotide probe I (DHS I) design 5'-p-(HEG)-p-GGAAGTCCAGCCCCATGGATGATGG where HEG-the rest of hexamethyleneimine, performed on an automatic DNA/RNA synthesizer ASM-800 using standard solid-phase hospitaliano method.

3. Receiving the modified electrode I

Spend the immobilization of the probe of earth I on the surface of carbon nanotubes. For this purpose, the working electrode modified with carbon nanotubes,placed in 10 ^-5M aqueous solution of DHS 1 so that the array of carbon nanotubes was completely immersed in the solution. Immobilization of the oligonucleotide to the surface of carbon nanotubes is carried out with stirring (speed 600 rpm) at 37°C for 18 of the Czech Republic. The electrode is dried for 2 h at 1-2 mm Hg in a desiccator with P₂O₅. The capacity of the nanotubes on the surface of the electrode relative to the oligonucleotide probe of earth I was 94 µmol/g

4. Detection of RNA using modified electrode I

To measure the electrochemical characteristics of the electrode by the method of cyclic voltammetry with lineinyi scan potential in three-electrode cell (figure 1) on the base (1) put the current collector auxiliary electrode (2), an auxiliary electrode placed two layers of separator (5) from non-woven polypropylene fibers. The separator excessively impregnated with 50 μl of the electrolyte solution, the electrode (3) and working electrode (4) with an array of carbon nanotubes concern separator (5). The composition of the electrolyte 0.1 M NaCl, 10 mm cacodylate sodium, 1 mm Na₂EDTA, pH 7.4. The electrodes connect to a recording device (6). Cyclic voltamperometry in the potential window for measurements [-0.2; 1] register using a potentiostat Elins R-30S, working in potentiodynamic mode. The potential is measured relative to silver chloride reference electrode. The scan speed capacity is 20 mV/cregistered capacity of the modified electrode in each cycle of operation of the electrode. Once equilibrium has been established on the separator put 50 μl of the solution of the RNA (5'-CUGCCAUCAUCCA0GGGGCUGGACUUCCUCU) in the same buffer. In the same mode register cyclic voltamperometry and calculate the capacity of the electrode. After the introduction of the sample see the change in the shape of a circular voltamperes corresponding to a hybridization probe the DHS I and detectable RNA and make a conclusion about the presence or absence in the sample RNA section, complementary to the oligonucleotide probe, and evaluate his concentration.

A typical example No. 2.

1. Receiving the modified working electrode

Modified working electrode receive as described in example No. 1.

2. Synthesis of oligonucleotide probe of earth II

Obtaining oligonucleotide probe II (DHS II) construction of 5'-Pyr-p-HEG-p-GGAAGTCCAGCCCCATGGATGATGG, where Pyr - balance pyrene, HEG - the rest of hexamethyleneimine, carried out by attaching piridiletilen to the 5'-terminal phosphate group of earth I by the method described in [Novopashin, DS, Totsky, O.S, Kholodar, S.A., Meshchaninov, M.I., Veniaminova, A.G. Bioorg. Chemistry. 2008. T. No. 5. S-682].

3. Receipt is modified electrode II

Spend the immobilization of the probe of earth II on the surface of carbon nanotubes as described in example No. 1. The capacity of the nanotubes on the surface of the electrode relative to the oligonucleotide probe of earth II was 157 µmol/g

4. Detection of RNA using modified electrode

Conduct measurements electrochemical characteristics of the electrode by the method of cyclic voltammetry with linear scan of potential in three-electrode cell as described in example No. 1. After introduction of the solution sample RNA (5'-CUGCCAUCAUCCAUGGGGCUGGACUUCCUCU) observe the change in the shape of a circular voltamperes: change their areas from cycle to cycle (figure 3) and the appearance of a peak (figure 4). The increase in capacity after adding a solution of detectable RNA is a signal that in the studied sample RNA contains a section, complementary to the immobilized oligonucleotide probe. The intensity appears on voltamperometry peak is proportional to the concentration of the detected RNA and is determined by the previously constructed calibration curve.

A typical example of No. 3

1. Receiving the modified working electrode

Modified working electrode receive as described in example No. 1.

2. Synthesis of oligonucleotide probe of earth III

Obtaining oligonucleotide probe III (DHS III) the con is e.g. the 5'-Pyr-pHEG-p-G ^mG^mA^mA^mG^mU^mC^mC^mA^mG^mC^mC^mC^mC^mA^mU^mG^mG^mA^mU^mG^mA^mU^mG^mG^mwhere Pyr - balance pyrene, HEG - the rest of hexamethyleneimine, G^m, A^mU^mand C^m- 2'-O-methylribonucleotide, carried out by attaching piridiletilen to the terminal phosphate group of the 5'-pHEG-p-G^mG^mA^mG^mU^mC^mC^mA^mG^mC^mC^mC^mC^mA^mU^mG^mG^mA^mU^mG^mA^mU^mG^mG^maccording to the method described in [Novopashin, D.S., Totsky, O.S, Kholodar, S.A., Meshchaninov, M.I., Veniaminova, A.G. Bioorg. Chemistry. 2008. T. No. 5. S-682]. The original synthesis of the oligonucleotide 5'-pHEG-p-

G^mG^mA^mA^mG^mU^mC^mC^mA^mG^mC^mC^mC^mC^mA^mU^mG^mG^mA^mU^mG^mA^mU^mG^mG^mperformed on an automatic DNA/RNA synthesizer ASM-800 standard solid-phase vospitannym method.

3. Receiving the modified electrode III

Spend immobilization probe the DHS III on the surface of carbon nanotubes as described in example No. 1. The capacity of the nanotubes on the surface of the electrode relative to the oligo is nucleotide probe amounted to 148 µmol/g

4. Detection of RNA using modified electrode III

Conduct measurements electrochemical characteristics of the electrode by the method of cyclic voltammetry with linear scan of potential in three-electrode cell as described in example No. 1. After introduction of the solution sample RNA (5'-CUGCCAUCAUCCAUGGGGCUGGACUUCCUCU) observe the change in the shape of a circular voltamperes: change their areas from cycle to cycle and the appearance of a peak, similar to that described in example No. 2. The increase in capacity after adding a solution of RNA is a signal that in the studied sample RNA contains a section, complementary to the immobilized oligonucleotide probe. The intensity appears on voltamperometry peak is proportional to the concentration of the detected RNA and is determined by the previously constructed calibration curve.

A typical example No. 4

1. Receiving the modified working electrode

Modified working electrode receive as described in example No. 1.

2. Synthesis of oligonucleotide probe of earth IV

Obtaining oligonucleotide probe IV (DHS IV) design 5-Pyr-R-AAAAAAAAAA, where Pyr - balance pyrene, carried out by attaching piridiletilen to the terminal phosphate group of the 5'-p-AAAAAAAAAA according to the method described in [Novopashin, D.S., Totsky, OS, Kholodar, With. the., Meshchaninov, M.I., Veniaminova, A.G. Bioorg. Chemistry. 2008. T. No. 5. S-682]. The synthesis of the original oligoribonucleotide 5'-p-AAAAAAAAAA performed on an automatic DNA/RNA synthesizer ASM-800 standard solid-phase vospitannym method.

3. Receiving the modified electrode IV

Spend immobilization probe the DHS SI on the surface of carbon nanotubes as described in example No. 1. The capacity of the nanotubes on the surface of the electrode relative to the oligonucleotide probe was 128* ámol/g

4. Detection of RNA using modified electrode IV

Conduct measurements electrochemical characteristics of the electrode

the method of cyclic voltammetry with linear scan of potential in three-electrode cell as described in example No. 1. After introduction of the solution sample RNA (5'-CCCCCCCC) see minor changes in the shape of cyclic voltamperometry. Build the dependence of the relative change in capacity from the number of the cycle (figure 5). See minor changes (up to 5%) capacity after adding solution. Make a conclusion about the absence in the sample RNA plot, a complementary oligonucleotide probe

Thus, the inventive method and device extend the capabilities of the method of detection of specific sequences NK (RNA or DNA) by registering elec is rahimizadeh characteristics of the electrodes by the method of cyclic voltammetry, simplify, cheapen and expedite it, allow the detection of the samples in small volumes, which is very essential in research and development of diagnostic systems. Furthermore, the method allows not only to detect a specific sequence NK (RNA or DNA), but also to estimate its concentration.

1. The method of detection of specific sequences of nucleic acids by electrochemical method, characterized in that the detection is carried out by registration cyclic voltamperometry working electrode modified with carbon nanotubes with ecovalence immobilized on their surface oligonucleotide probe, before and after inclusion in the study sample solution of nucleic acid (RNA or DNA); change in the capacitive characteristics defined by the area of cyclic voltamperometry, make a conclusion about the presence or absence in the sample of nucleic acid (RNA or DNA) of the site, a complementary oligonucleotide probe.

2. The method of detection according to claim 1, characterized in that after the registration of cyclic voltamperometry working electrode before and after inclusion in the study sample solution of nucleic acid (RNA or DNA) to build a profile of changes in the relative capacity of the electrode from the rooms of the loop.

3. Method detection specificationvalue nucleic acids by electrochemical method, characterized in that the detection is carried out by registration cyclic voltamperometry working electrode modified with carbon nanotubes with ecovalence immobilized on the surface of carbon nanotubes oligonucleotide probe containing the remainder of polyaromatic hydrocarbons as anchor groups for immobilization of the probe, before and after inclusion in the study sample solution of nucleic acid (RNA or DNA); change in the capacitive characteristics defined by the area of cyclic voltamperometry, or by the presence or absence of a peak on the capacitive characteristics make a conclusion about the presence or absence in the sample of nucleic acid (RNA or DNA) of the site, complementary to the oligonucleotide probe, and determine the concentration of detectable NK.

4. The method of detection according to claim 3, characterized in that after the registration of cyclic voltamperometry working electrode before and after inclusion in the study sample solution of nucleic acid (RNA or DNA) to build a profile of changes in the relative capacity of the electrode from the rooms of the loop.

5. The method according to claim 3, characterized in that the oligonucleotide probe modify the remnants of pyrene or other polyaromatic hydrocarbons.

6. The device for implementing the method of detection of specific sequences is kleinova acids by electrochemical method, representing electrochemical analyzer, consisting of a three-electrode electrochemical cell, the electrodes of which are connected to a recording device containing a power source, such as a potentiostat, characterized in that the working electrode is made of conductive silicon or metal substrate with grown therein vertically oriented carbon nanotubes, on the entire surface of which ecovalence immobilized oligonucleotide probe, complementary to the area defined NC, or modified oligonucleotide probe, complementary to the area defined NC; between the working and auxiliary electrodes is separator.

7. The device according to claim 6, characterized in that the oligonucleotide probe pre-modify polyaromatic hydrocarbons, which acts as an anchor group for non-covalent immobilization of the modified oligonucleotide probe on the surface of the carbon nanotubes of the working electrode.

8. The device according to claim 6, characterized in that the oligonucleotide probe modify the remnants of pyrene or other polyaromatic hydrocarbons.