CN102262122B - Single-wall carbon nano tube-based ultrasensitive deoxyribonucleic acid (DNA) biosensor and preparation method and application thereof - Google Patents
Single-wall carbon nano tube-based ultrasensitive deoxyribonucleic acid (DNA) biosensor and preparation method and application thereof Download PDFInfo
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- CN102262122B CN102262122B CN201110169043.9A CN201110169043A CN102262122B CN 102262122 B CN102262122 B CN 102262122B CN 201110169043 A CN201110169043 A CN 201110169043A CN 102262122 B CN102262122 B CN 102262122B
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Abstract
The invention provides a single-wall carbon nano tube-based ultrasensitive deoxyribonucleic acid (DNA) biosensor and a preparation method and application thereof. In the method, single-wall carbon nano tubes (SWCNTs) are grown on the surface of a silicon wafer on site by a chemical vapor deposition method, and gold nano particles are deposited on the surfaces of carbon nano tube electrodes by an electrochemical deposition technology. A single-stranded DNA (ssDNA) probe is self-assembled to the surfaces of SWCNTs-Au electrodes and subjected to hybridization reaction with complementary ssDNA. The change of electron transfer resistance before and after hybridization is recorded by utilizing the unique specific surface areas of the carbon nano tubes and the quick dynamic characteristics of the electrodes and by an electrochemical impedance method under the action of current signal amplification of nanometer gold to realize the quantitative detection of the complementary DNA. The detection limit of the sensor on the target DNA can reach between 10 and 20 M, so the sensor has the advantages of high sensitivity and selectivity, capability of being used repeatedly and the like, and has important significance in fields of medical diagnosis, the food industry, environment friendliness and the like.
Description
Technical field
The present invention relates to bioelectrochemical sensor and preparation field thereof, particularly a kind of DNA (deoxyribonucleic acid) (DNA) electrochemical sensor and preparation method thereof is applied with detecting.
Background technology
Gene diagnosis is disease to be made to the method for diagnosis by the existence of direct gene detection or defect.The detection object of gene diagnosis is DNA (deoxyribonucleic acid) or RNA (ribonucleic acid) (RNA).Aspect the methodology of gene diagnosis, successively set up restriction mapping analysis, making nucleic acid molecular hybridization, restriction fragment length polymorphism linkage analysis, PCR (PCR), and the DNA sensor that gets up of development in recent years and DNA chip technology etc.Labelling method nucleic acid hybridization detection technique now extensively can be used in again the relevant fields such as biology, medical science and environmental science, but its checkout procedure is time-consuming, effort, and traditional labelled with radioisotope poor stability, be difficult to meet the needs of each side, Development of Novel molecular hyridization Fast Detection Technique is extremely urgent.The DNA sensor provides a new way for the nucleic acid hybridization fast detecting, and it is to take the specificity of crossover process to be the quick sensing detection technology on basis.
Since human genome cracks, the development of DNA sensor field is maked rapid progress, and has obtained at present sizable progress.In each subjects such as bio-science, computer science, science, microelectronics, multiple theory and technology is used widely in this field.DNA sensor and DNA chip application are in the field of DNA sequencing, sudden change detection, genescreen, gene diagnosis and nearly all application nucleic acid hybridization.In numerous DNA sensor detecting methods, the electrochemical techniques based on the molecule electrical properties have unique superiority.Electrochemical measuring technique is sensitive, quick, cost is very cheap, and pick-up unit is light, low energy consumption and be easy to microminiaturizedly and integrated, meets the requirement of Handheld detection device and following chip lab (lab-on-a-chip).And the electrochemical gene sensing technology has following characteristics usually: hybridization reaction directly completes in its surface, and the change transitions that converter can produce crossover process becomes electric signal: according to change in electric amount before and after hybridization, thereby infer the amount of tested DNA; There is the superiority such as accurate, quick and cheap; The more important thing is, it can be promoted the use of in all fields relevant with DNA sequence dna information.
At present, synthetic and research and development application facet about nano material is rapid.The introducing of nano material makes the DNA sensor all greatly improve sensitivity and the selectivity of hybridization check.Carbon nano-tube (CNTs) is a kind of widely used bio-sensing base material, and golden nanometer particle is a kind of widely used antigen, antibody and cell marker, professor Mirkin of nanotechnology research institute of the Northwestern Univ USA leader's (Science of seminar, 1997,277,1078) utilizing oligonucleotides-modified golden nanometer particle to carry out having carried out pioneering research aspect the identification of DNA base.Variation by system color before and after hybridization realizes that the detection of DNA is had to the characteristics such as instrument is simple, experiment is convenient.But, due to the limitation of visual discrimination, make the sensitivity of aberration identification lower, so this method has certain limitation.Afterwards, this seminar has developed again the amplification method (Science of a kind of bio-bar-code of being called, 2003,301,1884), the method adopts the DNA-AuNP nano particle as signal probe, adopt magnetic-particle to connect capture probe simultaneously, the specific target sequence that captures from blending agent, form sandwich sandwich structure by DNA hybrid dna-AuNP nano particle, then the DNA of nanometer Au particle surface separated to detection.This method is very sensitive, but could detect after DNA need being separated, and has increased the step detected.For overcoming above-mentioned defect, the Fan Chun of Shanghai applied physics research institute sea teach problem group (CN 101245387A) combines enzymatic reaction and nm of gold enlarge-effect, in order to detect DNA, found that not only without DNA is separated to detection from the nm of gold surface, and kept sensitivity preferably (lowest detection is limited to the pM level) and selectivity.Yet regrettably the method need be introduced specific substrate, need to produce fluorescence or color is arranged after enzymic catalytic reaction, increase its cost, and limited to a certain extent its range of application.
Summary of the invention
The objective of the invention is the shortcoming and defect existed in order to overcome prior art, and a kind of sensitivity and specificity is higher and easy to detect, testing cost is low a kind of hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube are provided.
Another object of the present invention provides a kind of preparation method of the above-mentioned hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube.
In addition, the present invention also provides a kind of application of the hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube, the i.e. detection method of a kind of DNA.
For realizing first purpose of the present invention, technical scheme of the present invention is to include golden nanometer particle and as the Single Walled Carbon Nanotube of electrode basement, described golden nanometer particle is deposited on Single Walled Carbon Nanotube and forms Single Walled Carbon Nanotube/golden nanometer particle hybrid electrode, on this Single Walled Carbon Nanotube/golden nanometer particle hybrid electrode, is assembled with the ssDNA as the sulfydryl modification of probe.
Further arrange is that described Single Walled Carbon Nanotube for forming by the growth of chemical vapour deposition technique in-situ horizontal on silicon chip.
Further arranging is that described Single Walled Carbon Nanotube is the array structure that horizontal parallel is arranged, and array density is controlled at every 100 μ m intervals approximately 4-50 root Single Walled Carbon Nanotube, and the length of every Single Walled Carbon Nanotube is positioned at 100 μ m-2 mm.
Further arranging is that described golden nanometer particle pattern can be the golden dendrite with multilevel hierarchy.
The preparation method of the above-mentioned hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube comprises the following steps:
(1) the chemical vapor deposition growth Single Walled Carbon Nanotube is passed through in the growth of Single Walled Carbon Nanotube on silicon chip;
(2) with electrochemical method controllable deposition golden nanometer particle on the SWCNTs of growth, preparation Single Walled Carbon Nanotube/golden nanometer particle hybrid electrode;
(3) self assembly of electrode surface DNA probe, the probe ssDNA that sulfydryl (SH-) is modified carries out self assembly with the SWCNTs-Au electrode prepared, and obtains the hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube.
Further arranging is that described step (1) is to utilize the method that is rapidly heated, and with Fe/Mo, makes catalyzer, with ethanol, makes the carbon source for growth Single Walled Carbon Nanotube.
Further arranging is that described step (2) adopts two electrode systems, with platinum filament, does electrode, adopts chronoamperometry, and voltage is-0.1V--0.5v that sedimentation time is 5-50s, the HAuCl that concentration is 0.1-10 mM
4for Jin Yuan, the deposited gold nano particle.The further setting of this setting is that the technological parameter of described step (2) is that voltage is-0.5v, and sedimentation time is 8-15s.
Further arranging is that the self assembly condition of step (3) comprises electrode is immersed in the Tris-HCl buffer solution of the probe ssDNA modified containing the sulfydryl (SH-) of 1-μ M, and is placed in about 2-10 h under 1-10 ℃.
The present invention also provides the DNA detection method of the hypersensitive DNA biology sensor of a kind of utilization based on Single Walled Carbon Nanotube, its technical scheme: include following operation:
(1) hybridization, hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube will be placed in to DNA solution to be detected, at 35 ℃-45 ℃ hybridization 2h, then wash non-specific adsorption to the DNA that Single Walled Carbon Nanotube/do not hybridize on golden nanometer particle hybrid electrode surface;
(2) with electrochemical assay, at electrolytic solution, detect by the variation of the electrochemical parameter on the Single Walled Carbon Nanotube before and after step (1) hybridization/golden nanometer particle hybrid electrode surface, quantitatively or/and the concentration of qualitative detection DNA to be detected.
Further arranging is the electrochemical assay employing Electrode with Electrochemical Impedance Spectroscopy of described step (2), and frequency is 100 mHz-10kHz, and voltage is 0.17 V.
In the present invention, the said SWCNTs-Au electrode of having assembled the ssDNA of sulfydryl modification is to utilize golden nanometer particle and the DNA with the sulfydryl functional group interact and form.The English mark that wherein SWCNTs is Single Walled Carbon Nanotube, the English that SWCNTs-Au is Single Walled Carbon Nanotube/golden nanometer particle hybrid electrode is write a Chinese character in simplified form.
Complementary DNA(DNA probe or DNA to be detected in the present invention, also claim target dna) sulfydryl modification can adopt any prior art or commercial sources to obtain.
And the electrolytic solution of preparation method's step of the present invention in 4. can be electrolytic solution commonly used, as 1mM[Fe (CN)
4]
3-/4-; And electrochemical detection method used can be existing conventional electrochemical detection method, as electrochemical impedance or pulse voltammetry, the method for the preferred impedance of the present invention, quantitatively the electronics on detecting electrode surface transmits resistance.
The present invention utilizes gold and is modified with the interaction between the DNA of mercapto groups, and DNA probe is fixed on to the SWCNTs-Au electrode surface, by the hybridization of DNA molecular, catches the DNA with the DNA probe matching sequence.Because carbon nano-tube has capacitive character and specific surface area preferably, and the density of electrode surface carbon pipe can be controlled the density of golden nanometer particle, and then can control the DNA of self assembly and the DNA quantity of hybridization, effectively prevent the interference of winding between the DNA chain and signal conduction.Due to the amplification of golden nanometer particle, this sensor also has obvious signal when target sequence concentration is extremely low.The advantage of synthesise various, DNA sensor of the present invention can reach 10 to the detectability of object matching DNA
-20m, and have the ability of very strong differentiation single base mismatch, can reach 0.1pM to the detectability of single base mismatch DNA.SsDNA functionalization SWCNTs-Au modified electrode of the present invention has reusable advantage simultaneously.
Below in conjunction with specification drawings and specific embodiments, the present invention is described further.
The accompanying drawing explanation
Preparation and the measuring principle schematic diagram of Fig. 1 electrochemical DNA hybridization of the present invention sensor;
Fig. 2 is scanning electron microscope (SEM) figure of SWCNTs-Au electrode in one embodiment of the invention;
The SEM figure that Fig. 2 a is SWCNTs; The SEM figure that Fig. 2 b is the SWCNTs-Au that obtains after the depositing nano gold on SWCNTs; Fig. 2 c is the SWCNTs-Au partial enlarged drawing;
Fig. 3 (A) is that sensor of the present invention detects the electrochemical impedance figure that variable concentrations mates ssDNA fully; Curve a is that SWCNTs-Au is containing 1 mM [Fe (CN)
4]
3-/4-with the electrochemical impedance line in the Tris-HCl damping fluid of 0.1 M KCl; The b line is that after self assembly ssDNA, the SWCNTs-Au electrode is containing 1 mM [Fe (CN) 4]
3-/4-with the impedance line in the Tris-HCl damping fluid of 0.1 M KCl, curve c-m be respectively with contain 0.01 aM, 0.05 aM, 0.09 aM, 0.13 aM, 0.17 aM, 1.17 aM, 11.2 aM, 1 fM, 1 pM, 1 nM and 0.1 μ M coupling DNA hybridization after, the sensor is containing 1 mM [Fe (CN)
4]
3-/4-with the electrochemical impedance line in the Tris-HCl damping fluid of 0.1 M KCl.The mimic channel figure that illustration in Fig. 3 (A) is this working sensor;
Fig. 3 (B) is the SWCNTs-Au electrode of ssDNA functionalization and the relation that the electronics before and after the hybridization of variable concentrations target dna transmits increased resistance value and target dna concentration, and the electronics that the illustration in Fig. 3 (B) is this sensor transmits the relation curve of resistance change value and target dna molecule number;
Fig. 4 (A) adopts the single base mismatch that DNA electrochemical sensor of the present invention is implemented to detect.The a line is ssDNA(10 base) the SWCNTs-Au electrode of functionalization is containing 1 mM [Fe (CN)
4]
3-/4-transmit resistance with the electronics in the Tris-HCl damping fluid of 0.1 M KCl; The b line is that the electronics after the target dna of the sensor and tri-base mispairings of 0.1 pM is hybridized transmits resistance; The c line is that the electronics after the target dna of the sensor and 0.1 pM single base mismatch is hybridized transmits resistance; The d line is that the electronics after the sensor and the 0.1 pM target dna hybridization of mating fully transmits resistance, Fig. 4 (B) be adopt that DNA electrochemical sensor of the present invention implements respectively with the DNA hybridization of the single base mismatch of variable concentrations after, electronics transmits the linear relationship chart between the concentration of the changing value of resistance and single base mismatch DNA;
Fig. 5 is the relation curve that the electronics after electrochemical sensing electrode used in the present invention is hybridized with the 0.01aM DNA mated fully transmits resistance and regeneration times.
embodiment
Below by embodiment, the present invention is specifically described; only be used to further illustrate the present invention; can not be interpreted as limiting the scope of the present invention, the technician in this field can make some nonessential improvement and adjustment to the present invention according to the content of foregoing invention.
preferred embodiment
Preparation method of the present invention comprises the following steps:
(1) growth of SWCNTs
The utilization method that is rapidly heated, make catalyzer with Fe/Mo, with ethanol, makes carbon source for growth SWCNTs;
(2) preparation of SWCNTs-Au electrode adopts two electrode systems, with platinum filament, does electrode, and SWCNTs is working electrode, and voltage is-0.5v that sedimentation time is 10s, HAuCl
4for golden source deposited gold nano particle, then with MilliQ, clean.
(3) assembling of electrode surface DNA probe (1) hybridization is placed in the electrode made in step (3) DNA solution of coupling, at 42 ℃ of hybridization 2h, then with MilliQ, cleans.Wash the coupling DNA that non-specific adsorption is not hybridized to electrode surface.
(2) Electrochemical Detection is at 1 mM [Fe (CN)
4]
3-/4-transmit resistance, the quantitative concentration that detects coupling DNA with the electronics on Electrode with Electrochemical Impedance Spectroscopy detection assembling front and back and hybridization front and back electrode surface in (containing 0.1M KCl).
Below by following several embodiment and experimental data, further set forth ability and the high sensitivity of differentiation single base mismatch of the present invention.
(1) preparation of SWCNTs-Au electrode
Adopt the chemical vapor deposition (CVD) method as shown in Fig. 1, at first with heating furnace, quartz ampoule is rapidly heated to 1000 ℃, then passes into ethanol/helium mix gas, with the Fe/Mo nano particle, make catalyzer, high temperature reduction reacts 15 min, in silica-based situ growth, obtains single-wall carbon nanotube array.Then electrode is connected as working electrode to the HAuCl of 10ml (1mM) with copper wire
4for Jin Yuan, using the Pt silk as to electrode, adopt the method for constant potential, and regulation voltage is-0.5V that, with after two electrode system deposition 10s, powered-down, use the MilliQ water wash, dries up electrode surface with ear washing bulb and obtain the SWCNTs-Au electrode.
(2) preparation of sensing electrode and hybridization
The DNA sequence dna of selecting in the present embodiment is as follows:
Probe ssDNA:5 '-SH-CCC CAT CCC C-3 '
Mate DNA:5 '-GGG GAT GGG G-3 ' fully
The SWCNTs-Au electrode obtained in (1) is immersed in the ssDNA solution of 1 μ M of about 1ml to self assembly 10h under the condition of 4 ℃.Take out and repeatedly to rinse electrode surface with MilliQ water, dry up and characterize with impedance afterwards;
The sensing electrode of gained is put into to the coupling DNA solution that concentration is 0.1 μ M, under 45 ℃, hybridization 2h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.
(3) recording electrochemical impedance curve (EIS) is the electrode immersion solution of making 1mM[Fe (CN)
4]
3-/4-in the electrolytic solution of (containing 0.1MKCl), then carry out the electrochemical impedance experiment, obtain the EIS curve, the electronics that records sensing electrode after the circuit matching transmits resistance (as curve m in Fig. 2 A).
Obtain by the method in embodiment 1 the coupling DNA solution that sensing electrode is put into concentration 0.1 μ M, under 45 ℃, hybridization 10h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.Record the electrochemical impedance curve by the method in embodiment 1, result is consistent with embodiment 1.
The method of pressing in embodiment 1 obtains sensing electrode, the coupling DNA solution that to put into concentration be 1nM, under 45 ℃, hybridization 2h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.Record electrochemical impedance curve (as curve l in Fig. 2 A) by the method in embodiment 1.
The method of pressing in embodiment 1 obtains sensing electrode, the coupling DNA solution that to put into concentration be 1pM, under 45 ℃, hybridization 2h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.Record electrochemical impedance curve (as curve k in Fig. 2 A) by the method in embodiment 1.
The method of pressing in embodiment 1 obtains sensing electrode, the coupling DNA solution that to put into concentration be 1fM, under 45 ℃, hybridization 2h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.Record electrochemical impedance curve (as curve j in Fig. 2 A) by the method in embodiment 1.
embodiment 6
The method of pressing in embodiment 1 obtains sensing electrode, the coupling DNA solution that to put into concentration be 1.17aM, under 45 ℃, hybridization 2h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.Record electrochemical impedance curve (as curve h in Fig. 2 A) by the method in embodiment 1.
embodiment 7
The method of pressing in embodiment 1 obtains sensing electrode, the coupling DNA solution that to put into concentration be 0.13aM, under 45 ℃, hybridization 2h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.Record electrochemical impedance curve (as curve f in Fig. 2 A) by the method in embodiment 1.
embodiment 7
The method of pressing in embodiment 1 obtains sensing electrode, the coupling DNA solution that to put into concentration be 0.01aM, under 45 ℃, hybridization 2h forms SWCNTs-Au-dsDNA, take out and repeatedly rinse electrode surface with MilliQ water and Tris-HCl buffer solution, wash away the DNA that non-specific adsorption is not hybridized at electrode surface.Record electrochemical impedance curve (as curve c in Fig. 2 A) by the method in embodiment 1.
The result (Fig. 2) that to sum up embodiment obtains is known: one, DNA probe has obtained effectively fixing on SWCNTs-Au; Two, the hybridization reaction of the sensing electrode of probe and target DNA can make EIS obtain the significant variation of resistance generation.Thereby this bio-sensing electrode has or not target dna sequence in judgement sample solution qualitatively.The least concentration that can measure 10 base sequences by the method is 0.01aM.
By the method in Application Example 1, on target DNA, exist the sequence of 3 base mispairings and 1 base mispairing as target dna, least concentration all is located at 0.1pM.As shown in Figure 5, when 0.1pM, 3 base mispairings and strand sensor do not have significant change to result, with resistance and the strand sensor of gained after the aim sequence hybridization of 1 base of mispairing, a little increase are arranged, and with taget DNA, significant resistance difference are arranged.Visible, sensor of the present invention has the ability of very strong detection single base mismatch, and sequence-specific is very high.The DNA sequence dna adopted in the present embodiment is:
1-?Mismatched:?5’-GGG?GTT?GGG?G-3’
3-Mismatched:?5’-GGG?GCC?CGG?G-3’
embodiment 9
By the sensing electrode after target DNA in step (2) in embodiment 1 and 0.1 μ M hybridization in hot water in sex change 10min after, with the MilliQ water of ice, rinse rapidly, again in hot water after sex change 10min, after rinsing and dry up with the MilliQ water of ice rapidly again, recross, 4 circulations of sex change.Probe groups loading amount and hybridization efficiency remain unchanged substantially, and as shown in Figure 5, by the high-temperature denatured process of several circulations, the acquisition performance of regeneration electrode remains unchanged result substantially as seen.
Claims (9)
1. the hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube, it is characterized in that: include golden nanometer particle and as the Single Walled Carbon Nanotube of electrode basement, described golden nanometer particle is deposited on Single Walled Carbon Nanotube and forms Single Walled Carbon Nanotube/golden nanometer particle hybrid electrode, be assembled with the ssDNA as the sulfydryl modification of probe on this Single Walled Carbon Nanotube/golden nanometer particle hybrid electrode, described Single Walled Carbon Nanotube is the array structure that horizontal parallel is arranged, array density is controlled at every 100 μ m intervals approximately 4-50 root Single Walled Carbon Nanotube, the length of every Single Walled Carbon Nanotube is positioned at 100 μ m-2 mm.
2. a kind of hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube according to claim 1, it is characterized in that: a kind of as substrate in silicon chip, quartz glass or sapphire of described Single Walled Carbon Nanotube, and form by the growth of chemical vapour deposition technique in-situ horizontal.
3. a kind of hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube according to claim 1, it is characterized in that: described golden nanometer particle pattern can be the golden dendrite with multilevel hierarchy.
4. the preparation method of the hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube as claimed in claim 1 is characterized in that comprising the following steps:
(1) the chemical vapor deposition growth Single Walled Carbon Nanotube is passed through in the growth of Single Walled Carbon Nanotube on silicon chip;
(2) with electrochemical method controllable deposition golden nanometer particle on the SWCNTs of growth, preparation Single Walled Carbon Nanotube/golden nanometer particle hybrid electrode;
(3) self assembly of electrode surface DNA probe, the probe ssDNA that sulfydryl (SH-) is modified carries out self assembly with the Single Walled Carbon Nanotube prepared/golden nanometer particle hybrid electrode, obtains the hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube.
5. the preparation method of a kind of hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube according to claim 4 is characterized in that: described step (1) adopts Fe/Mo to make catalyzer, with ethanol, makes the carbon source for growth Single Walled Carbon Nanotube.
6. the preparation method of a kind of hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube according to claim 5, it is characterized in that: described step (2) adopts two electrode systems, with platinum filament, do electrode, adopt chronoamperometry, voltage is-0.1V--0.5v, sedimentation time is 5-50s, the HAuCl that concentration is 0.1-10 mmol/L
4for Jin Yuan, the deposited gold nano particle.
7. the preparation method of a kind of hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube according to claim 6, it is characterized in that: the self assembly condition of step (3) comprises electrode is immersed in the Tris-HCl buffer solution of the probe ssDNA modified containing the sulfydryl (SH-) of 1 ~ 10 μ mol/L, and is placed in about 2-10 h under 1-10 ℃.
8. a DNA detection method of utilizing the hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube claimed in claim 1 is characterized in that: include following operation:
(1) hybridization, hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube is placed in to DNA solution to be detected, at 35 ℃-45 ℃ hybridization 2-10 h, then wash non-specific adsorption to the DNA that Single Walled Carbon Nanotube/do not hybridize on golden nanometer particle hybrid electrode surface;
(2) with electrochemical assay, in electrolytic solution, detect by the variation of the electrochemical parameter on the Single Walled Carbon Nanotube before and after step (1) hybridization/golden nanometer particle hybrid electrode surface, quantitatively or/and the concentration of qualitative detection DNA to be detected.
9. a kind of DNA detection method of utilizing the hypersensitive DNA biology sensor based on Single Walled Carbon Nanotube claimed in claim 1 according to claim 8, it is characterized in that: the electrochemical assay of described step (2) adopts Electrode with Electrochemical Impedance Spectroscopy, frequency is 100 mHz-10kHz, and voltage is 0.17 V.
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| "基于多壁碳纳米管和纳米金修饰电极测定特殊序列DNA 的电化学生物传感器的制备";何月珍等;《分析试验室》;20110131;第30卷(第1期);第20页第2栏倒数第1段-第21页第1栏第3段 * |
| 何月珍等."基于多壁碳纳米管和纳米金修饰电极测定特殊序列DNA 的电化学生物传感器的制备".《分析试验室》.2011,第30卷(第1期), |
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