CN1885036A - Photo-electro-chemical method for detecting nucleic acid - Google Patents
Photo-electro-chemical method for detecting nucleic acid Download PDFInfo
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- CN1885036A CN1885036A CN 200510011991 CN200510011991A CN1885036A CN 1885036 A CN1885036 A CN 1885036A CN 200510011991 CN200510011991 CN 200510011991 CN 200510011991 A CN200510011991 A CN 200510011991A CN 1885036 A CN1885036 A CN 1885036A
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Abstract
The invention relates to a method for detecting nucleic acid alkali group with optical electric chemical technique, wherein said method comprises: a, contacting the tested sample and the material with optical electric chemical activity; b, evaluating the oxidization and reduction reaction between the nucleic acid alkali group and said optical electric chemical material, that using light to transform the optical electric chemical material into exciting state with existed electrode, and processing oxidization reduction reaction with nucleic acid alkali group when the excited material loses electrons, to evaluate the current generated in oxidization reduction reaction; c, based on said current, analyzing the sample qualitative or quantitatively. The invention can be used in biological detection, as nucleic acid cross detection, nucleic hurt detection and chemical material gene toxicity detection.
Description
Technical field
The present invention relates to the detection technique field of nucleic acid, particularly relate to the detection nucleic acid hybridization of Optical Electro-Chemistry qualitative, quantitative and the method for damage.
Technical background
Know that now human diseases all have direct or indirect relation with gene, therefore, method for quick highly sensitive, high selectivity to nucleic acid exists more and more urgent demand, and the monitoring of biological pollutant all has realistic meaning in this toxicity screening for gene studies, drug screening, clinical diagnosis and ecological toxicology research, noval chemical compound, the environment.
Many detection methods have been used to detect nucleic acid, for example, present detection of nucleic acids southern blotting technique, RNA trace and round pcr commonly used, a common ground in the operation of these methods be must the probe of not hybridization and primer with could detect target nucleic acid by other signals after crossbred separates, the complex operation complexity, time-consuming, technical difficulty is big, and is difficult to nucleic acid is carried out accurate detection by quantitative.
Nucleic acid hybridization be set up the earliest, the most basic nucleic acid detection technique, this analytical approach generally need detect target nucleic acid attached to the label on the target nucleic acid chain, what these labels adopted the earliest is radioactive nuclide, after biotin, fluorescein are arranged.Radioisotope labeling is highly sensitive, yet relatively more expensive, dangerous.Biotin and fluorescein are safe in utilization, but sensitivity is relatively poor, especially can't carry out sensitive detection to the damage of base in the nucleic acid and the mispairing of single base-pair, for example: United States Patent (USP) 5,143,854 and 5,405,783.
The catalytic action that utilizes nucleic acid base is directly detected the existing description of nucleic acid, for example, United States Patent (USP) 6,361,951 have described and have a kind ofly directly detected the method for nucleic acid hybridization with cyclic voltammetry, and this method is with the base catalysis current signal of nucleic acid, detect nucleic acid hybridization according to the intensity of catalytic current.Although described method can directly detect the redox reaction of nucleic acid base, do not need mark, the sensing range of this method is limited, and this method comes the excitation signal molecule with voltage on electrode, and the range of choice of signaling molecule is restricted.
Summary of the invention
The objective of the invention is to overcome the deficiencies in the prior art, provide a kind of easy, effectively, Optical Electro-Chemistry detects the method for nucleic acid efficiently.
For achieving the above object, technical solution of the present invention provides the method that a kind of Optical Electro-Chemistry detects nucleic acid base, comprising:
(a) testing sample is contacted with the material with Optical Electro-Chemistry activity under appropriate condition;
(b) redox reaction between above-mentioned sample amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up the Optical Electro-Chemistry active substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the electric current that redox reaction produced;
(c), sample is carried out qualitative or quantitative test according to the electric current that redox reaction produced.
Described method, the analyte in its described testing sample is nucleotide, oligonucleotides or nucleic acid.
Described method, the nucleic acid base in its described sample are guanine, adenine, 6-thioguanine, 8-oxidation guanine or 8-oxoadenine compound.
Described method, its described Optical Electro-Chemistry active substance are inorganic molecule, organic molecule, inorganic/organic compound molecule, inorganic polymer, organic polymer, inorganic/the organic composite polymeric thing, inorganic particulate matter, organic particulate matter or inorganic/organic composite granulated thing.
Described method, its described Optical Electro-Chemistry active substance, its material are metal/organic ligand complex compound.
Described method, its described metal/organic ligand complex compound, its metal is ruthenium or osmium.
Described method, its described metal/organic ligand complex compound, its organic ligand is nitrogenous heteroaromatic.
Described method, its described organic ligand be dipyridine, connect the phenanthrol of the ter cycloheptapyridine of company's pyrazine of the dipyridine of pyrazine, ter cycloheptapyridine, phenanthrol, phthalein cyanogen dyestuff, replacement, replacement, replacement, replacement, two pyridos [3,2-a; 2 ', 3 '-c] azophenlyene or adjacent two single aza-phenanthrenes.
Described method, its described electrode are metal electrode, nonmetal electrode or metal oxide electrode.
Described method, its described metal oxide electrode material is tin ash.
Described method, its described metal oxide electrode is to be prepared on conductive matrices by the nano particle tin ash.
Described method, testing sample in its described (a) step comprises the nucleic acid samples of nucleic acid hybridization sample or damaged.
Described method, the Optical Electro-Chemistry detection method of its described nucleic acid hybridization sample comprises:
(a) the nucleic acid hybridization sample is contacted with the material with Optical Electro-Chemistry activity under appropriate condition;
(b) variation of the redox reaction between above-mentioned sample amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up photoelectric chemical substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the variation of the hybridization front and back electric current that redox reaction produced;
(c) change according to the electric current that redox reaction produced before and after the hybridization, sample is carried out qualitative or quantitative test.
Described method, the Optical Electro-Chemistry detection method of the nucleic acid samples of its described damaged comprises:
(a) nucleic acid samples with damaged contacts with the material with Optical Electro-Chemistry activity under appropriate condition;
(b) redox reaction between above-mentioned sample amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up photoelectric chemical substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the variation of the nucleic acid damaging front and back electric current that redox reaction produced;
(c) according to the variation of the electric current that redox reaction produced before and after the nucleic acid damaging, sample is carried out qualitative or quantitative test.
Described method, the nucleic acid samples of its described damaged is the damage that chemical factor causes, for the caused base of alkylating reagent comes off, the base alkylation, the DNA bound rupture, the DNA chain is crosslinked, or nitrite and base form the damage that diazo salt causes, or intercalator is because of inserting the damage that the DNA chain causes, or replace normal base to participate in the DNA chain disturbing the synthetic damage that causes of dna replication dna owing to base analogue enters cell, or because synthetic, the chemical substance that exists in the environment is because of changing dna sequence dna, base on the modifying DNA chain, influence the caused damage of dna replication dna.
Described method, the nucleic acid samples of its described damaged are the damages that physical factor causes, the damage that causes for ultraviolet ray, high-energy ray or ionising radiation;
Described method, the damage that its described dna molecular oneself factor causes is the spontaneous chemical change of mistake in the dna replication dna or DNA.
Described method is used for the detection of chemical substance genotoxicity, and it comprises:
(a) chemical substance to be measured and nucleic acid probe are carried out injury response;
(b) above-mentioned reaction product is contacted with the material with Optical Electro-Chemistry activity under appropriate condition;
(c) redox reaction between above-mentioned product amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up photoelectric chemical substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the variation of the nucleic acid damaging front and back electric current that redox reaction produced;
(d), the genotoxicity of described chemical substance is carried out qualitative or quantitative test according to the variation of the electric current that redox reaction produced before and after the nucleic acid damaging.
Described method, its described chemical substance is environmental contaminants or its metabolin.
Described method, its described environmental contaminants are palycyclic aromatic or its metabolin.
Described method, its described using up changes into excited state with photoelectric chemical substance, and its illuminating source is: the cathode modulation of hollow, xenon arc lamp, xenon-mercury lamp, metal halide lamp, light emitting diode or laser.
Description of drawings
Fig. 1 is the photocurrent that is adsorbed on the ruthenium two (2,2 '-two pyridines) (4 '-methyl-4-carboxyl-2,2 '-two pyridines) on the tin oxide nano electrode.
Fig. 2 is the spectrum that is adsorbed on the ruthenium two (2,2 '-two pyridines) (4 '-methyl-4-carboxyl-2,2 '-two pyridines) on the nano tin dioxide semi-conducting electrode.
Fig. 3 is the acid catalyzed photocurrent of variable concentrations guanosine.
Fig. 4 detects the nucleic acid damaging schematic diagram for Optical Electro-Chemistry.
Fig. 5 detects the photocurrent of damage dna for Optical Electro-Chemistry.
Fig. 6 is the photocurrent of variable concentrations damage dna.
Embodiment
The invention provides the method that a kind of Optical Electro-Chemistry detects nucleic acid base, comprising: (a) testing sample is contacted with the material with Optical Electro-Chemistry activity under appropriate condition; (b) redox reaction between above-mentioned sample amplifying nucleic acid base of evaluation and the described Optical Electro-Chemistry active substance is carried out qualitative or quantitative test to sample; Described be evaluated as under the situation that electrode exists, to use up the Optical Electro-Chemistry active substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the electric current that redox reaction produced.
In the method for the present invention, when having the material of Optical Electro-Chemistry activity with optical excitation, the ground state electronics can transit to excited state L (E) from ground state L (G) because of absorbing photon energy, as shown in Equation 1.
Optical excitation: L (G)+hv----→ L (E)
(1)
The electronics that is in excited state has very strong activity, can lose easily, migrates on the electrode that has than low energy level, produces the bias light electric current, as shown in Equation 2.
Photocurrent produces: L (E)+electrode----→ L (O)+electrode (e)
(2)
Photoelectric chemical substance becomes oxidation state L (O) because of losing an electronics.The redox-potential of the particular bases guanine (G) of nucleic acid, adenine (A), 6-thioguanine, 8-oxo-guanine and 8-oxo-adenine is lower, can be used as reductive agent and uses.If there are these bases in the solution, photoelectric chemical substance will be got back to original reduction ground state, and enter into the redox reaction of a new round, as shown in Equation 3, like this these particular bases with regard to catalysis photo-signal.
Photocurrent catalysis: L (O)+particular bases----→ L (G) (3)
Different its catalytic capabilities with base sequence of the structure of nucleic acid molecules are also just different, and therefore, detected in the above final photocurrent intensity is also just different, and this just provides theoretical foundation for the qualitative and quantitative detection of nucleic acid.
The invention provides a kind of Optical Electro-Chemistry pick-up unit, comprising: (a) sample cell, the wall energy in this pond enough see through the light of exciting light electrochemical substance; (b) electrode is fit to analyze the electric current that electron transfer produced between photoelectric chemical substance and the electrode; (c) has the photoelectric chemical substance of photoelectric activity; (d) light source can the exciting light electrochemical substance; (e) redox reaction detector, photo-signal that can the analyzing and testing base catalysis.
Arbitrary shape can need be processed into by various concrete experiments in described nucleic acid samples pond.
Described electrode is metal electrode, nonmetal electrode or metal oxide electrode, and electric current can enter or leave medium by them.
Described photoelectric chemical substance is that described Optical Electro-Chemistry active substance is inorganic molecule, organic molecule, inorganic/organic compound molecule, inorganic polymer, organic polymer, inorganic/the organic composite polymeric thing, inorganic particulate matter, organic particulate matter or inorganic/organic composite granulated thing.Described photoelectric chemical substance has following feature: stronger absorption is arranged in visible wavelength region; The energy level of excited state is higher than the energy level of electrode, guarantees that electron transfer can take place; The life-span long enough of excited state, it is luminous that electron transfer is better than; The oxidation state of photoelectric chemical substance is stable with going back ortho states.
Described photoelectric chemical substance can be fixed on the electrode, mark is to above-mentioned nucleic acid or be dissolved in the medium.Described using up changes into excited state with photoelectric chemical substance, and its light emitting source is cathode modulation, xenon arc lamp, xenon-mercury lamp, metal halide lamp, light emitting diode or the laser of hollow.
Test objective according to different can arbitrarily design concrete implementation step of the present invention.Such as, testing goal can be: the A. Optical Electro-Chemistry detects nucleic acid hybridization; B. Optical Electro-Chemistry detects nucleic acid damaging; C. Optical Electro-Chemistry detects chemical substance genotoxicity etc.
Device before detecting is prepared:
1. the preparation of nano-stannic oxide semi-conducting electrode
Indium tin oxide-coated glass is used washing agent (15 minutes), pure water (2 minutes respectively, twice), acetone (5 minutes), isopropyl alcohol (5 minutes) and pure water (10 minutes, twice) supersound washing, dry up, the tin ash colloidal solution uniform spreading of getting the 15nm particle diameter is on the ITO battery lead plate.After the drying, 450 ℃ of sintering were cut into the small electrode sheet of 0.5cm * 2.5cm after 1 hour, promptly obtained the nano-stannic oxide semi-conducting electrode.
2. the preparation of ruthenium complex mark avidin
(concentration is 10mg/mL with 2mg avidin avidin, volume is 200uL, solvent is the phosphate buffer of 30mM, PH 8.0), drop into the anhydrous N that 30uL contains 0.7mg ruthenium two (2,2 '-two pyridines) (4 '-methyl-4-carboxyl-2,2 '-two pyridines)-succinamide ester, in the N-dimethylformamide solution, and shake reaction 90 minutes in the dark.Ruthenium complex on unmarked is gone out through centrifuging, must be through the good biotin protein of ruthenium complex mark (Ru-Avidin).The BCA method is adopted in the concentration determination of albumen, and the content of ruthenium is measured its absorption value at the 470nm place with ultraviolet-visible pectrophotometer.The albumen that mark is good is dissolved in 20mM phosphate buffer PH 7.3, is made into the solution of 0.2mg/mL.
3.Ru-Avidin decorated nanometer tin ash semi-conducting electrode
Getting the nanometer titanium dioxide tin electrode that the above-mentioned Ru-Avidin solution of 10uL uniform spreading prepares in the foregoing description 2 adsorbs, contact area is 0.5 square centimeter, adsorbs after 1 hour, and water shakes washes 5 minutes, airing promptly obtains to modify good semiconductor working electrode.
Photocurrent is measured
Under time-based pattern, measuring light electric current on CHI 630 electrochemical analysers.Light source is the laser of the 470nm wavelength of a 20mW.The low-light electrochemical cell is to be made by the glass of polishing, and contrast electrode is Ag/AgCl, is platinum electrode to electrode.Light beam is vertically injected in the pond, acts on stannic oxide electrode at its back side.Manual mobile lighttight cardboard makes light beam intermittent irradiation electrode clocklike.The ruthenium bipyridyl complexes has absorption maximum at 470nm.
As shown in Figure 1, the nano oxidized tin electrode that has adsorbed Ru-Avidin is placed in the photoelectrochemistrpool pool in the phosphate buffer of 20mM, and PH 5.5.With cardboard light beam is covered when measuring beginning, open cardboard being taken out every 10 seconds.The complex compound of photoelectric chemical substance ruthenium can produce photocurrent under the exciting of light as can be seen, but because there is not catalyzer to exist, photocurrent is unsettled, along with the consumption of ruthenium complex, and the decline that photocurrent continues.
As shown in Figure 2, the nano oxidized tin electrode that has adsorbed Ru-Avidin is placed in the photoelectrochemistrpool pool of phosphate buffer of 20mM, and PH 5.5.Light source is the xenon lamp of a 500W, and the wavelength with the monochromator selection needs forwards the monochromator selector switch between 450nm and the 650nm to, and detects corresponding photocurrent.The spectrogram that obtains like this is similar to the absorption spectrum of ruthenium, shows that photocurrent is the generation that excites by ruthenium complex.
Embodiment 2
The detection of guanine catalysis photocurrent
Phosphate buffer PH 5.5 with 20mM is diluted to following several concentration gradient with guanylic acid: 1 μ g/mL, 5 μ g/mL, 10 μ g/mL, 20 μ g/mL and 50 μ g/mL.Carry out Photoelectric Detection in the guanylic acid solution adding photoelectrochemistrpool pool that dilution is good, get the acid catalyzed photocurrent of variable concentrations guanosine.(a is 0 μ g/mL to the result as shown in Figure 3; B is 1 μ g/mL; C is 5 μ g/mL; D is 10 μ g/mL; E is 20 μ g/mL; F is 50 μ g/mL).This experimental result shows that the base guanine has very strong catalytic action to the photocurrent that photoelectric chemical substance produces, and the high more catalytic action of its concentration is also just strong more.
The dna damage experiment
Use styrene oxide as damage reagent, styrene oxide is the metabolic product that styrene produces after the cytochrome P 450 enzymes catalysis in liver, styrene is a kind of important industrial chemical, because distributing and extensively be present in the environment in the Industrial Engineering is a kind of ubiquitous toxic pollutant in the environment.The injury response of calf androgenic gland DNA carries out in 20mM phosphate buffer PH 5.5, and temperature of reaction is 37 ℃, and the reaction time is 30 minutes, so obtains the DNA through the styrene oxide damage.Styrene oxide is a carcinogen, and experimental implementation should be carried out with closed container in fuming cupboard as far as possible, prevents to suck or the human body contact.
Embodiment 4
Optical Electro-Chemistry detects dna damage
The principle of Optical Electro-Chemistry detection dna damage as shown in Figure 4; dna damage rear section base is oxidized; the destruction that causes the dna double chain structure, double-stranded protection base is down reacted its participation that is more prone to by discharging in various degree; bring into play its catalytic action; therefore, compare, can strengthen photosignal greatly with unmarred DNA; the intensity of different its photosignals of the degree of dna damage is also just different, and therefore this photoelectrochemical system can the sensitive DNA that detects damage.
Concrete test experience: the nano oxidized tin electrode that a slice has been adsorbed Ru-Avidin is positioned in the photoelectrochemistrpool pool, get 600uL, PH 5.5, the phosphate buffer of 20mM joins in the photoelectrochemistrpool pool, transfer laser intensity to 10mW, obtain the bias light electric current of phosphate buffer under without any material catalysis.Get unmarred calf androgenic gland DNA (CT-DNA) solution (600uL of 0.05mg/mL, be dissolved in the phosphate buffer of 20mM, PH 5.5) join in the photoelectrochemistrpool pool, open laser, the photocurrent of CT-DNA, the photocurrent of CT-DNA is better than the background current value, and DNA is described, and reaction has catalytic action to Optical Electro-Chemistry.(concentration is 0.05mg/mL through the CT-DNA solution of embodiment 6 steps damage to get 600uL; be dissolved in the phosphate buffer of 20mM; PH 5.5) join in the electrolytic cell, open laser, the photocurrent of the CT-DNA that must damage; not damaging the photocurrent of CT-DNA A compares; the damage CT-DNA the photoelectricity flow valuve further strengthen, illustrate that DNA is damaged after, its duplex structure is destroyed; double-stranded protection base down by in various degree discharge the catalytic action enhancing.(a is the phosphoric acid background to the result as shown in Figure 5; B is the photocurrent that does not damage CT-DNA; C is the photocurrent of damage CT-DNA; Laser every 10 seconds switches once).
Embodiment 5
Optical Electro-Chemistry detects the damage dna of variable concentrations
Phosphate buffer PH 5.5 with 20mM is made into following several concentration gradient with the CT-DNA that damages: 0.15mg/mL, 0.1mg/mL, 0.05mg/mL, 0.02mg/mL and 0.005mg/mL.The CT-DNA solution of 5 kinds of damages that prepare is added in the photoelectrochemistrpool pool, carry out Photoelectric Detection according to the mode of embodiment 7, get the photocurrent of variable concentrations damage CT-DNA, (a is 0.15mg/mL to the result as shown in Figure 6; B is 0.1mg/mL; C is 0.05mg/mL; D is 0.02mg/mL; E is 0.005mg/mL; F is 0mg/mL).This experimental result shows, photoelectrochemical system is to the detection sensitivity very of damage dna, even when its concentration has only 0.005mg/mL, compares its photoelectricity flow valuve with background current and still has clearly enhancing.
Claims (21)
1, a kind of Optical Electro-Chemistry detects the method for nucleic acid base, it is characterized in that, comprising:
(a) testing sample is contacted with the material with Optical Electro-Chemistry activity;
(b) redox reaction between above-mentioned sample amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up the Optical Electro-Chemistry active substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the electric current that redox reaction produced;
(c), sample is carried out qualitative or quantitative test according to the electric current that redox reaction produced.
2. method according to claim 1 is characterized in that, the analyte in the described testing sample is nucleotide, oligonucleotides or nucleic acid.
3. method according to claim 1 is characterized in that, the nucleic acid base in the described sample is guanine, adenine, 6-thioguanine, 8-oxidation guanine or 8-oxoadenine compound.
4. method according to claim 1, it is characterized in that described Optical Electro-Chemistry active substance is inorganic molecule, organic molecule, inorganic/organic compound molecule, inorganic polymer, organic polymer, inorganic/the organic composite polymeric thing, inorganic particulate matter, organic particulate matter or inorganic/organic composite granulated thing.
5. method according to claim 4 is characterized in that, described Optical Electro-Chemistry active substance, its material are metal/organic ligand complex compound.
6. method according to claim 5 is characterized in that, described metal/organic ligand complex compound, its metal are ruthenium or osmium.
7. method according to claim 5 is characterized in that, described metal/organic ligand complex compound, its organic ligand are nitrogenous heteroaromatic.
8. method according to claim 7, it is characterized in that, described organic ligand is dipyridine, connect the phenanthrol of the ter cycloheptapyridine of company's pyrazine of the dipyridine of pyrazine, ter cycloheptapyridine, phenanthrol, phthalein cyanogen dyestuff, replacement, replacement, replacement, replacement, two pyridos [3,2-a; 2 ', 3 '-c] azophenlyene or adjacent two single aza-phenanthrenes.
9. method according to claim 1 is characterized in that, described electrode is metal electrode, nonmetal electrode or metal oxide electrode.
10. method according to claim 9 is characterized in that, described metal oxide electrode material is tin ash.
11. method according to claim 10 is characterized in that, described metal oxide electrode is to be prepared on conductive matrices by the nano particle tin ash.
12. method according to claim 1 is characterized in that, testing sample in described (a) step comprises the nucleic acid samples of nucleic acid hybridization sample or damaged.
13. method according to claim 12 is characterized in that, the Optical Electro-Chemistry detection method of described nucleic acid hybridization sample comprises:
(a) the nucleic acid hybridization sample is contacted with the material with Optical Electro-Chemistry activity;
(b) variation of the redox reaction between above-mentioned sample amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up photoelectric chemical substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the variation of the hybridization front and back electric current that redox reaction produced;
(c) change according to the electric current that redox reaction produced before and after the hybridization, sample is carried out qualitative or quantitative test.
14. method according to claim 12 is characterized in that, the Optical Electro-Chemistry detection method of the nucleic acid samples of described damaged comprises:
(a) nucleic acid samples with damaged contacts with the material with Optical Electro-Chemistry activity;
(b) redox reaction between above-mentioned sample amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up photoelectric chemical substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the variation of the nucleic acid damaging front and back electric current that redox reaction produced;
(c) according to the variation of the electric current that redox reaction produced before and after the nucleic acid damaging, sample is carried out qualitative or quantitative test.
15. method according to claim 14, it is characterized in that, the nucleic acid samples of described damaged is the damage that chemical factor causes, for the caused base of alkylating reagent comes off, the base alkylation, the DNA bound rupture, the DNA chain is crosslinked, or nitrite and base form the damage that diazo salt causes, or intercalator is because of inserting the damage that the DNA chain causes, or replace normal base to participate in the DNA chain disturbing the synthetic damage that causes of dna replication dna owing to base analogue enters cell, or because synthetic, the chemical substance that exists in the environment is because of changing dna sequence dna, base on the modifying DNA chain, influence the caused damage of dna replication dna.
16. method according to claim 14 is characterized in that, the nucleic acid samples of described damaged is the damage that physical factor causes, the damage that causes for ultraviolet ray, high-energy ray or ionising radiation.
17. method according to claim 14 is characterized in that, the damage that described dna molecular oneself factor causes is the spontaneous chemical change of mistake in the dna replication dna or DNA.
18. method according to claim 1 is used for the detection of chemical substance genotoxicity, it is characterized in that, comprising:
(a) chemical substance to be measured and nucleic acid probe are carried out injury response;
(b) above-mentioned reaction product is contacted with the material with Optical Electro-Chemistry activity;
(c) redox reaction between above-mentioned product amplifying nucleic acid base of evaluation and the described photoelectric chemical substance, promptly under the situation that electrode exists, use up photoelectric chemical substance is changed into excited state, after the material that is excited loses electronics, carry out redox reaction with nucleic acid base, estimate the variation of the nucleic acid damaging front and back electric current that redox reaction produced;
(d), the genotoxicity of described chemical substance is carried out qualitative or quantitative test according to the variation of the electric current that redox reaction produced before and after the nucleic acid damaging.
19. method according to claim 18 is characterized in that, described chemical substance is environmental contaminants or its metabolin.
20. method according to claim 19 is characterized in that, described environmental contaminants are palycyclic aromatic or its metabolin.
21. according to claim 1,13,14 or 18 described methods, it is characterized in that, described using up changes into excited state with photoelectric chemical substance, and its illuminating source is: the cathode modulation of hollow, xenon arc lamp, xenon-mercury lamp, metal halide lamp, light emitting diode or laser.
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| CN102539741B (en) * | 2010-10-29 | 2014-12-24 | 希森美康株式会社 | Detection device for detecting a test substance, electrode substrate, working electrode, inspection tip, method of detecting a test substance |
| CN102539741A (en) * | 2010-10-29 | 2012-07-04 | 希森美康株式会社 | Detection device for detecting a test substance, electrode substrate, working electrode, inspection tip, method of detecting a test substance, and method of detecting a sample substance |
| CN103649741B (en) * | 2011-07-04 | 2016-11-16 | 日本电气方案创新株式会社 | Evaluate the method for the redox active of nucleic acid molecules and there is the nucleic acid molecules of redox active |
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| US9637737B2 (en) | 2011-07-04 | 2017-05-02 | Nec Solution Innovators, Ltd. | Method for evaluating redox activity of nucleic acid molecule and nucleic acid molecule having redox activity |
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| US10138480B2 (en) | 2011-07-04 | 2018-11-27 | Nec Solution Innovators, Ltd. | Method for evaluating redox activity of nucleic acid molecule and nucleic acid molecule having redox activity |
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| CN103048392A (en) * | 2011-10-13 | 2013-04-17 | 蔡剑平 | Kit and detection system for evaluating aging degree of mammal and application of 8-oxoG |
| CN104535762A (en) * | 2015-01-15 | 2015-04-22 | 上海市杨浦区中心医院 | Method for efficiently screening out DNA chain scission injury protective drugs |
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| CN106222266A (en) * | 2016-07-29 | 2016-12-14 | 中国科学院生态环境研究中心 | A kind of method detecting nucleic acid base disappearance and dedicated kit thereof |
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