CN102253092B - Composite film modified DNA sensor and its preparation method and application in detection of lignin peroxidase (Lip) specific coding gene segment - Google Patents
Composite film modified DNA sensor and its preparation method and application in detection of lignin peroxidase (Lip) specific coding gene segment Download PDFInfo
- Publication number
- CN102253092B CN102253092B CN 201110097748 CN201110097748A CN102253092B CN 102253092 B CN102253092 B CN 102253092B CN 201110097748 CN201110097748 CN 201110097748 CN 201110097748 A CN201110097748 A CN 201110097748A CN 102253092 B CN102253092 B CN 102253092B
- Authority
- CN
- China
- Prior art keywords
- carbon paste
- carbon
- electrode
- paste electrode
- dna sensor
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Abstract
The invention discloses a composite film modified DNA sensor. The DNA sensor is characterized in that the sensor comprises a carbon paste electrode; the carbon paste electrode comprises a carbon rod, a Teflon tube, a magnet, and carbon paste; an induction end of the carbon paste electrode is coated with a sensitive substance; the sensitive substance comprises a composite film and a DNA capture probe; magnetic nanoparticles, multi-walled carbon nanoscale tube-gold nanoparticles and chitosan are utilized for modifying orderly the surface of the carbon paste electrode and compose the composite film; and the DNA capture probe is utilized for modifying the surface of the multi-walled carbon nanoscale tube-gold nanoparticles. The invention also discloses a preparation method of the DNA sensor. The preparation method comprises the processing steps of manufacturing a carbon paste electrode, modifying the surface of an induction end of the manufactured carbon paste electrode by a sensitive substance, and the like. The invention further discloses an application of the DNA sensor in detecting a lignin peroxidase (Lip) specific coding gene segment. Because a sensitive substance is utilized for modifying the surface of a carbon paste electrode of the DNA sensor, the DNA sensor has a strong capacity of electron conduction and a high accuracy of detection. The preparation method has the advantages of low cost and simple process.
Description
Technical field
The present invention relates to field of biosensors, relate in particular to a kind of process biology sensor after film modified and its preparation method and application.
Background technology
Adopting biosensor technique to detect genetic fragment is a trend of gene analysis test, and wherein electrochemical sensor is paid close attention to widely, highly sensitive because it possesses response fast, high selectivity and the advantage such as workable.For the DNA sensor, common working electrode is gold electrode or based on the screen printing electrode of nm of gold, because crosslinked by sulfydryl and gold of the gene probe that is modified with sulfydryl is easy to be assemblied in electrode surface, carries out coherent detection.But, gold electrode is expensive, and the screen printing electrode is disposable electrode, and for a large amount of detections, its total cost is also very considerable.In recent years, along with novel nano-material and the swift and violent combination of electrochemical sensing technology, a class is developed rapidly based on the electrochemical sensor of paramagnetism technique for fixing.This class sensor is filled into carbon paste in teflon (PTFE) pipe, and in the carbon paste of magnet intercalation electrode front portion, be formed on the carbon paste electrode that the electrode front-end surface has fixed magnetic field, and then, the recycling paramagnetism attracts the magnetic nanoparticle of fixed function, makes up the specific detection that different sensors is used for the related objective thing.From the operating cost angle, adopt economic magnetic nanoparticle and carbon paste electrode to replace above-mentioned two class electrodes to carry out genetic fragment and detect a kind of feasible way of can yet be regarded as.But have a problem can not be ignored: the electronic conduction ability of carbon paste electrode is starkly lower than gold electrode, can reduce the accuracy of detection of gene sensing technology.
In order to improve the electronic conduction ability of carbon paste electrode, in the sensor construction strategy, learn relevant feature in conjunction with new material, use some nano materials and biomolecule material to form structure of composite membrane and be modified at the carbon paste electrode surface.Nano material, because have high electronic conductivity, can provide bigger serface such as carbon nano-tube, magnetic nano-particle and golden nanometer particle etc., keep the advantages such as biologically active, be considered to outstanding biomolecule carrier and signal and transmit medium, can improve the sensitivity of electrochemical sensor.Except nano material, a kind of biomolecule material, shitosan is strong because of nontoxic, biological fitness, have the characteristics such as film forming ability also is widely used in the structure of biology sensor.
Summary of the invention
Technical matters to be solved by this invention is: for the deficiencies in the prior art, provide the complex film modified DNA sensor that a kind of cost is low, electronic conduction ability is strong, accuracy of detection is high, also corresponding preparation method and the application of this DNA sensor in detecting lignin peroxidase (Lip) specific coding genetic fragment that a kind of this DNA sensor is provided.
For solving the problems of the technologies described above, the present invention by the following technical solutions:
A kind of complex film modified DNA sensor, comprise a carbon paste electrode, described carbon paste electrode comprises carbon-point and PTFE tube, described carbon-point is built in the described PTFE tube, described carbon-point one end is drawn described PTFE tube by electric wire, the described carbon-point other end contacts with a magnet, be filled with carbon paste between described magnet and the described PTFE tube mouth of pipe, the induction end of described carbon paste electrode is coated with sensitive materials, described sensitive materials comprise the DNA capture probe of composite membrane and sulfhydrylation, described composite membrane is by modifying successively in the magnetic nanoparticle of the sulfhydrylation on described carbon paste electrode surface, multi-walled carbon nano-tubes-the golden nanometer particle of sulfhydrylation, and the shitosan composition, described DNA capture probe is modified in described multi-walled carbon nano-tubes-golden nanometer particle surface.
As further improvement of these options, described magnet is arranged at apart from the mouth of pipe of described polyfluortetraethylene pipe and is not more than the 8mm place.
In the technique scheme, the sequence preference of described DNA capture probe is 5 '-HS (CH
2)
6TTGTTGACGAAGGACTGCCA-3 '.
As a total technical conceive, the present invention also provides a kind of preparation method of above-mentioned complex film modified DNA sensor, may further comprise the steps:
(1) makes carbon paste electrode: in PTFE tube, put into carbon-point, and insert magnet at described carbon-point one end, form magnetic regions, fill with carbon paste between described magnet and the PTFE tube mouth of pipe, the carbon-point other end is drawn PTFE tube by electric wire, obtain the crude green body of carbon paste electrode, more described crude green body is carried out surface treatment, obtain final carbon paste electrode;
(2) decorating carbon paste electrode surface: the induction end surface of the magnetic nanoparticle for preparing and multi-walled carbon nano-tubes-golden nanometer particle and chitosan solution being modified successively the carbon paste electrode that step (1) makes, naturally obtain complex film modified carbon paste electrode after drying, DNA capture probe with ready sulfhydrylation drops in through complex film modified carbon paste electrode induction end surface again, after the self assembly, obtain complex film modified DNA sensor.
The technological process of above-mentioned preparation magnetic nanoparticle is preferably: prepare Fe under nitrogen protection
3O
4Then gelatinous precipitate adds polyglycol, positive silane ethyl ester, methyl alcohol, aminopropyl trimethoxysilane and halfcystine successively, obtains the magnetic nanoparticle of sulfhydrylation after the reaction.
The technological process of above-mentioned preparation multi-walled carbon nano-tubes-golden nanometer particle is preferably: with the multi-walled carbon nano-tubes purifying, react with halfcystine again, make the multi-walled carbon nano-tubes sulfhydrylation, then with multi-walled carbon nano-tubes and the golden nanometer particle reaction of sulfhydrylation, obtain the multi-walled carbon nano-tubes-golden nanometer particle of sulfhydrylation.
The self assembly principle of the modification of the composite membrane of DNA sensor of the present invention and DNA capture probe as shown in Figure 2.
As a total inventive concept, the present invention also provide utilize above-mentioned complex film modified DNA sensor (sequence of DNA capture probe is 5 '-HS (CH
2)
6TTGTTGACGAAGGACTGCCA-3 ') method that Lip specific coding genetic fragment is detected may further comprise the steps:
(1) choose the sequence of object chain, and the sequence of modelled signal probe:
The sequence of object chain is 5 '-TGGCAGTCCTTCGTCAACAA-3 '; According to the sequence of object chain, the sequence of modelled signal probe is 5 '-Biotin-TGGCAGTCCTTCGTCAACAA-3 ';
(2) competitive hybridization reaction: described signal probe and solution to be measured are splashed into the induction end surface of described complex film modified DNA sensor, and competitive reaction is no less than 60 minutes;
(3) signalase amplifies: again pH is DNA sensor surface neutral, that phosphate buffered solution that contain Avidin-horseradish peroxidase (SA-HRP) drops in the competitive hybridization reaction, the lower reaction of physiological temp (for example 37 ℃) is no less than 30 minutes, forms signalase connection amplification system;
(4) enzymic catalytic reaction: at last with benzenediol and H
2O
2Be substrate, join the DNA sensor of amplification system as working electrode to contain described horseradish peroxidase (HRP) signalase, set up the electrolytic cell of three-electrode system, adopt the reduction current that produces in the chronoamperometry mensuration enzyme-catalyzed reaction to change, if the reduction current fall is not less than 6.9%, judge and contain lignin peroxidase specific coding genetic fragment in the solution to be measured, finish detection.
Above-mentioned detection method is based on following principle (as shown in Figure 3) and finishes: the HRP of mark on the signal probe, and can be at H
2O
2Catalytic decomposition p-dihydroxy-benzene under the condition that exists is when add p-dihydroxy-benzene and H in electrolytic cell
2O
2After, HRP makes a hydroxyl on the p-dihydroxy-benzene lose H
+Become the carbonyl of two keys, provide electronics to make its reduction by electrode surface again, this process can produce detectable response signal and reach identifying purpose, and according to the content of the strong and weak corresponding Lip specific coding genetic fragment of the signal that produces, thereby realize mensuration to Lip specific coding genetic fragment.
In the technique scheme, the reduction potential that detects in the described electrolytic cell is preferably-0.3V~-0.2V, most preferably be-0.252V, described electrolytic solution cell is preferably the neutral phosphate buffered solution of pH value.
In the technique scheme, if judge and contain lignin peroxidase specific coding genetic fragment in the solution to be measured, then can be according to the linear relationship between object chain mrna concentration and the galvanochemistry hybridization signal, set up equation of linear regression, measure the size of the concentration that contains lignin peroxidase specific coding genetic fragment in the solution to be measured, equation of linear regression can obtain by the following method:
Repeat the step of above-mentioned detection method, (2)~, (4), detect the hybridization solution of the Lip specific coding genetic fragment of a series of known variable concentrations, and according to the current-responsive result under each certain concentration, (referring to Fig. 4), can draw the linear relationship between object chain mrna concentration and the galvanochemistry hybridization signal, can set up equation of linear regression, by experiment, the linear equation that draws is: DP%=, (3.4908 ± 0.1725) x+, (29.8172 ± 0.6892), (seeing Fig. 5), wherein, DP% is the electric current suppression ratio
I0 is the catalytic reduction electric current that object chain has neither part nor lot in competitive hybridization when reaction HRP, I
xThe catalytic reduction electric current of HRP when participating in the competition hybridization reaction for object chain; X is the natural logarithm of object chain concentration, and the range of linearity of concentration is 0.001 μ molL
-1~1 μ molL
-1, be limited to 1nmolL under detecting
-1The foundation that Lip specific coding genetic fragment concentration was calculated after this equation of linear regression can be used as.
Compared with prior art, the invention has the advantages that: the present invention is in conjunction with new bio molecule and nano material, with nano materials and chitosan-modified in the carbon paste electrode surface such as carbon nano-tube, magnetic nano-particle and golden nanometer particles, make up electrochemical DNA biosensor, greatly strengthen the electronic conduction ability of carbon paste electrode, thereby improve accuracy of detection; The DNA sensor, method that preparation of the present invention is complex film modified, with low cost, technique is simple; Utilize complex film modified DNA sensor of the present invention that Lip specific coding genetic fragment is detected, efficient, the low-cost dynamic change situation that detects gene of energy, thus the monitoring and the control procedure that can be the microbial degradation lignin provide a kind of effective biology tool.
Description of drawings
Fig. 1 is the structural representation of complex film modified DNA sensor of the present invention;
Fig. 2 is the composite membrane of complex film modified DNA sensor of the present invention and the assembling synoptic diagram of capture probe;
Fig. 3 is the hybridization reaction schematic diagram of complex film modified DNA sensor of the present invention;
Fig. 4 is that the present invention makes up in the linear equation process with chronoamperometry and detects the electric current that the Lip specific coding genetic fragment of variable concentrations obtains-time changing curve figure.
Fig. 5 is the linear regression graph of Lip specific coding genetic fragment content of the present invention and curent change.
Marginal data:
1, electric wire; 2, PTFE tube; 3, carbon-point; 4, magnet; 5, carbon paste; 6, sensitive materials (structure of composite membrane and DNA capture probe).
Embodiment
The present invention is further illustrated below in conjunction with specific embodiment and accompanying drawing.
A kind of complex film modified DNA sensor as shown in Figure 1, comprise a carbon paste electrode, carbon paste electrode comprises carbon-point 3 and PTFE tube 2, carbon-point 3 is built in the PTFE tube 2, carbon-point 3 one ends are drawn PTFE tube 2 by electric wire 1, carbon-point 3 other ends contact with a magnet 4, magnet 4 is arranged at next-door neighbour's carbon-point 3 one ends and apart from the mouth of pipe 8mm place of PTFE tube 2, be filled with carbon paste 5 between magnet 4 and PTFE tube 2 mouths of pipe, the induction end of carbon paste electrode is coated with sensitive materials 6, sensitive materials 6 comprise composite membrane and sulfhydrylation the DNA capture probe (sequence is 5 '-HS (CH
2)
6TTGTTGACGAAGGACTGCCA-3 '), composite membrane by modify successively the magnetic nanoparticle in the sulfhydrylation on carbon paste electrode surface, multi-walled carbon nano-tubes-golden nanometer particle and the shitosan of sulfhydrylation forms, the DNA capture probe is modified in multi-walled carbon nano-tubes-golden nanometer particle surface.
A kind of preparation method of above-mentioned complex film modified DNA sensor comprises following concrete steps:
(1) makes carbon paste electrode: in PTFE tube, put into carbon-point, be close to carbon-point one end and put into magnet apart from PTFE tube mouth of pipe 8mm place, form magnetic regions, fill with carbon paste between magnet and the PTFE tube mouth of pipe, the carbon-point other end is drawn PTFE tube by electric wire, obtains the crude green body of carbon paste electrode, again with the surface finish of electrode crude green body, then water flushing electrode surface is used HNO more successively
3(massfraction is 50%), acetone, water carry out ultrasonic cleaning, again with the phosphate buffer flushing, naturally dry at last, obtain final carbon paste electrode;
(2) preparation magnetic nanoparticle and multi-walled carbon nano-tubes-golden nanometer particle composite membrane:
The technological process of preparation magnetic nanoparticle is: prepare Fe under nitrogen protection
3O
4Then gelatinous precipitate adds polyglycol, positive silane ethyl ester, methyl alcohol, aminopropyl trimethoxysilane and halfcystine successively, obtains the magnetic nanoparticle of sulfhydrylation after the reaction;
The technological process of preparation multi-walled carbon nano-tubes-golden nanometer particle is: use H
2O
2With sulfuric acid mixture liquid with the multi-walled carbon nano-tubes purifying, after the vacuum drying, with the reaction of itself and halfcystine, sulfydryl in modifications, multi-walled carbon nano-tubes and the golden nanometer particle with sulfhydrylation reacts again, obtains the multi-walled carbon nano-tubes-golden nanometer particle of sulfhydrylation
(3) decorating carbon paste electrode surface: the induction end surface of the magnetic nanoparticle for preparing and multi-walled carbon nano-tubes-golden nanometer particle and chitosan solution being modified successively the carbon paste electrode that step (1) makes, naturally obtain complex film modified carbon paste electrode after drying, again with the DNA capture probe of ready sulfhydrylation (sequence is 5 '-HS (CH
2)
6TTGTTGACGAAGGACTGCCA-3 ') drops in through complex film modified carbon paste electrode induction end surface, carry out self assembly by sulfydryl and the effect of gold, with the damping fluid flushing, obtain complex film modified DNA sensor after drying again.
The DNA sensor that utilizes above-described embodiment to make is 0.012 μ molL to concentration respectively
-1, 0.59 μ molL
-1With 0.22 μ molL
-1Lip specific coding genetic fragment testing sample measure, the detection of the Lip specific coding genetic fragment testing sample of each concentration is all carried out with the following step:
(1) choose object chain and modelled signal probe:
The sequence of choosing object chain is 5 '-TGGCAGTCCTTCGTCAACAA-3 '; According to the sequence of object chain, the sequence of modelled signal probe is 5 '-Biotin-TGGCAGTCCTTCGTCAACAA-3 ';
(2) competitive hybridization reaction: signal probe and object chain testing sample are splashed into respectively the electrode surface of the DNA sensor that contains capture probe, 37 ℃ of lower competitive reactions 60 minutes;
(3) signalase amplifies: the phosphate buffered solution (pH 6.98) that will contain again SA-HRP drops in the DNA sensor surface that reacts through competitive hybridization, and 37 ℃ of lower reactions 30 minutes form signalase connection amplification system;
(4) enzymic catalytic reaction: containing 1mmolL
-1Add 0.5mmolL in the phosphate buffer of p-dihydroxy-benzene (pH 7.38)
-1H
2O
2, with p-dihydroxy-benzene and H
2O
2Be substrate, join the DNA sensor of amplification system as working electrode to contain the HRP signalase, with saturated calomel electrode preferably as contrast electrode, with platinum plate electrode preferably as to electrode, set up the electrolytic cell of three-electrode system, adopt chronoamperometry, measure the DNA sensor under reduction potential-0.252V, the reduction current that HRP and substrate catalytic reaction produce changes that (electrochemical gaging is to adopt CHI660B electro-chemical systems that Shanghai occasion China instrument company produces to be connected with three-electrode system in the 50mL electrolytic cell, to control and to monitor), test result is as shown in table 1 below, by descending as seen from Table 1, the reduction current fall of three kinds of solution to be measured all is not less than 6.9%, can judge thus all to contain lignin peroxidase specific coding genetic fragment in three kinds of solution to be measured;
(5) quantitative test: according to reduction current fall measured in the following table 1, and in conjunction with equation of linear regression DP%=(3.4908 ± 0.1725) x+ (29.8172 ± 0.6892) that has made up, can determine Lip specific coding genetic fragment concentration value in three groups of testing samples, testing result sees the following form 1.
Table 1: the testing result of three kinds of variable concentrations testing samples
Testing sample μ molL -1 | Reduction current fall % | Measure concentration μ molL -1 | Recovery % |
0.012 | 14.4 | 0.013±0.001 | 105.8 |
0.59 | 27.6 | 0.608±0.010 | 102.1 |
0.22 | 24.7 | 0.212±0.01 | 97.4 |
The result can clearly be seen that from said determination, this method is easy and simple to handle, and is highly sensitive, and selectivity is good, can be efficiently, the low-cost online Lip specific coding genetic fragment content that detects, for monitoring and the control procedure of microbial degradation lignin provides technical support.
The above only is preferred implementation of the present invention, and protection scope of the present invention also not only is confined to above-described embodiment, and all technical schemes that belongs under the thinking of the present invention all belong to protection scope of the present invention.Should be pointed out that for those skilled in the art the some improvements and modifications not breaking away under the principle of the invention prerequisite all should be considered as protection scope of the present invention.
Claims (9)
1. complex film modified DNA sensor, comprise a carbon paste electrode, described carbon paste electrode comprises carbon-point and polyfluortetraethylene pipe, described carbon-point is built in the described polyfluortetraethylene pipe, described carbon-point one end is drawn described polyfluortetraethylene pipe by electric wire, the described carbon-point other end contacts with a magnet, be filled with carbon paste between described magnet and the described polyfluortetraethylene pipe mouth of pipe, the induction end of described carbon paste electrode is coated with sensitive materials, it is characterized in that: described sensitive materials comprise the DNA capture probe of composite membrane and sulfhydrylation, described composite membrane is by modifying successively in the magnetic nanoparticle of the sulfhydrylation on described carbon paste electrode surface, multi-walled carbon nano-tubes-the golden nanometer particle of sulfhydrylation, and the shitosan composition, described DNA capture probe is modified in described multi-walled carbon nano-tubes-golden nanometer particle surface;
The sequence of described DNA capture probe is 5 '-HS (CH
2)
6TTGTTGACGAAGGACTGCCA-3 '.
2. complex film modified DNA sensor according to claim 1 is characterized in that: described magnet is arranged at apart from the mouth of pipe of described polyfluortetraethylene pipe and is not more than the 8mm place.
3. the preparation method of a complex film modified DNA sensor as claimed in claim 1 or 2 is characterized in that, comprises following processing step:
(1) makes carbon paste electrode: in polyfluortetraethylene pipe, put into carbon-point, and insert magnet at described carbon-point one end, form magnetic regions, fill with carbon paste between described magnet and the polyfluortetraethylene pipe mouth of pipe, the carbon-point other end is drawn polyfluortetraethylene pipe by electric wire, obtain the crude green body of carbon paste electrode, more described crude green body is carried out surface treatment, obtain carbon paste electrode;
(2) decorating carbon paste electrode surface: the induction end surface of the magnetic nanoparticle for preparing and multi-walled carbon nano-tubes-golden nanometer particle and chitosan solution being modified successively the carbon paste electrode that step (1) makes, naturally obtain complex film modified carbon paste electrode after drying, DNA capture probe with ready sulfhydrylation drops in through complex film modified carbon paste electrode induction end surface again, after the self assembly, obtain complex film modified DNA sensor.
4. preparation method according to claim 3 is characterized in that, described magnetic nanoparticle is to prepare by following steps: prepare Fe under nitrogen protection
3O
4Then gelatinous precipitate adds polyglycol, positive silane ethyl ester, methyl alcohol, aminopropyl trimethoxysilane and halfcystine successively, obtains the magnetic nanoparticle of sulfhydrylation after the reaction.
5. preparation method according to claim 3, it is characterized in that, described multi-walled carbon nano-tubes-golden nanometer particle is to prepare by following steps: with the multi-walled carbon nano-tubes purifying, react with halfcystine again, make the multi-walled carbon nano-tubes sulfhydrylation, then with multi-walled carbon nano-tubes and the golden nanometer particle reaction of sulfhydrylation, obtain the multi-walled carbon nano-tubes-golden nanometer particle of sulfhydrylation.
6. one kind is utilized complex film modified DNA sensor as claimed in claim 1 to the method that lignin peroxidase specific coding genetic fragment detects, and may further comprise the steps:
(1) choose object chain and modelled signal probe:
The sequence of choosing object chain is 5 '-TGGCAGTCCTTCGTCAACAA-3 '; According to the sequence of object chain, the sequence of modelled signal probe is 5 '-Biotin-TGGCAGTCCTTCGTCAACAA-3 ';
(2) competitive hybridization reaction: described signal probe and solution to be measured are splashed into the induction end surface of described complex film modified DNA sensor, and competitive reaction is no less than 60 minutes;
(3) signalase amplifies: be neutrality and the phosphate buffered solution that contains Avidin-horseradish peroxidase again with pH, drop in the induction end surface of the reacted DNA sensor of competitive hybridization, reaction is no less than 30 minutes under the physiological temp, forms horseradish peroxidase signalase connection amplification system;
(4) enzymic catalytic reaction: at last with benzenediol and H
2O
2Be substrate, join the DNA sensor of amplification system as working electrode to contain described horseradish peroxidase signalase, set up the electrolytic cell of three-electrode system, adopt the reduction current that produces in the chronoamperometry mensuration enzyme-catalyzed reaction to change, if the reduction current fall is not less than 6.9%, judge and contain lignin peroxidase specific coding genetic fragment in the solution to be measured, finish detection.
7. method according to claim 6, it is characterized in that, if judge and contain lignin peroxidase specific coding genetic fragment in the solution to be measured, detect again the concentration of the genetic fragment of lignin peroxidase specific coding described in the solution to be measured according to following equation of linear regression;
Described equation of linear regression is: DP%=(3.4908 ± 0.1725) x+ (29.8172 ± 0.6892), and wherein, DP% is the electric current suppression ratio,
I
0The catalytic reduction electric current of horseradish peroxidase when having neither part nor lot in the competitive hybridization reaction for object chain, I
xThe catalytic reduction electric current of horseradish peroxidase when participating in the competition hybridization reaction for object chain; X is the natural logarithm of described lignin peroxidase specific coding genetic fragment concentration, and the range of linearity that concentration detects is 0.001 μ molL
-1~1 μ molL
-1, be limited to 1nmolL under detecting
-1
8. according to claim 6 or 7 described methods, it is characterized in that: the three-electrode system in the described step (4) be with saturated calomel electrode as contrast electrode, with platinum plate electrode as to electrode.
9. according to claim 6 or 7 described methods, it is characterized in that: the reduction potential that detects in the described electrolytic cell for-0.3V~-0.2V, electrolytic solution cell is that pH value is the phosphate buffered solution of neutrality.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 201110097748 CN102253092B (en) | 2011-04-19 | 2011-04-19 | Composite film modified DNA sensor and its preparation method and application in detection of lignin peroxidase (Lip) specific coding gene segment |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN 201110097748 CN102253092B (en) | 2011-04-19 | 2011-04-19 | Composite film modified DNA sensor and its preparation method and application in detection of lignin peroxidase (Lip) specific coding gene segment |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102253092A CN102253092A (en) | 2011-11-23 |
CN102253092B true CN102253092B (en) | 2013-05-29 |
Family
ID=44980474
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN 201110097748 Expired - Fee Related CN102253092B (en) | 2011-04-19 | 2011-04-19 | Composite film modified DNA sensor and its preparation method and application in detection of lignin peroxidase (Lip) specific coding gene segment |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102253092B (en) |
Families Citing this family (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103344689B (en) * | 2013-07-01 | 2015-08-12 | 北京工业大学 | Based on Fe 3o 4magnetic pole, the preparations and applicatio of-AuNPs magnetic composite nanoparticles immobilization acetylcholinesteraseelectrochemistry |
CN103743805B (en) * | 2014-01-17 | 2016-07-27 | 湖南大学 | Biosensor based on aza mesoporous carbon, preparation method and applications |
CN104267078B (en) * | 2014-10-09 | 2016-08-24 | 无锡百灵传感技术有限公司 | A kind of electrochemical sensor of quick detection tripolycyanamide |
CN105092683B (en) * | 2015-08-31 | 2018-02-23 | 湖南大学 | It is a kind of to be used to detect electrochemical sensor of lead and its preparation method and application |
CN105466986A (en) * | 2015-11-17 | 2016-04-06 | 中国科学院电子学研究所 | Preparation method of electrochemical biosensor sensitive membrane |
CN105466980B (en) * | 2015-11-23 | 2017-12-05 | 北京农业信息技术研究中心 | A kind of plant live body ascorbic acid detection method based on microelectrode biosensor |
CN105675676B (en) * | 2016-01-22 | 2018-04-13 | 湖南大学 | Electrochemical DNA biosensor for detecting mercury ion or cysteine and its preparation method and application |
CN107085025B (en) * | 2016-02-15 | 2019-09-17 | 太原师范学院 | A kind of self assembly trilamellar membrane electrode and its preparation method and application |
CN105758918B (en) * | 2016-04-08 | 2018-10-12 | 青岛科技大学 | A kind of preparation of the DNA sensor based on electroreduction graphene oxide and decorated by nano-gold electrode and application process |
CN106282344B (en) * | 2016-08-15 | 2018-03-02 | 广州市宝创生物技术有限公司 | Nano-Au probe function and service thing of high stable and preparation method thereof |
CN110376380B (en) * | 2019-07-25 | 2020-07-24 | 华中科技大学 | Electrochemical enzyme-linked immunosensor and preparation and application thereof to antigen detection |
CN115078497B (en) * | 2022-05-30 | 2023-07-14 | 宁德师范学院 | Amino carbon nano tube carbon paste electrode, preparation method and application |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101982764A (en) * | 2010-11-01 | 2011-03-02 | 湖南大学 | Composite membrane modified biosensor and preparation method and application thereof |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999067628A1 (en) * | 1998-06-24 | 1999-12-29 | Therasense, Inc. | Multi-sensor array for electrochemical recognition of nucleotide sequences and methods |
-
2011
- 2011-04-19 CN CN 201110097748 patent/CN102253092B/en not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101982764A (en) * | 2010-11-01 | 2011-03-02 | 湖南大学 | Composite membrane modified biosensor and preparation method and application thereof |
Non-Patent Citations (3)
Title |
---|
An electrochemical DNA sensor based on a layers–film construction modified electrode;Yi Zhang,Guang-Ming Zeng,Lin Tang, et al;《Analyst》;20110824;第136卷;4204-4210 * |
Yi Zhang,Guang-Ming Zeng,Lin Tang, et al.An electrochemical DNA sensor based on a layers–film construction modified electrode.《Analyst》.2011,第136卷4204-4210. |
章毅,曾光明,汤琳,等.基于磁性纳米粒子固定技术的漆酶传感器用于垃圾堆肥中邻苯二酚的检测.《环境科学》.2007,第28卷(第10期),2320-2325. * |
Also Published As
Publication number | Publication date |
---|---|
CN102253092A (en) | 2011-11-23 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102253092B (en) | Composite film modified DNA sensor and its preparation method and application in detection of lignin peroxidase (Lip) specific coding gene segment | |
Zhang et al. | A sensitive amperometric AChE-biosensor for organophosphate pesticides detection based on conjugated polymer and Ag-rGO-NH2 nanocomposite | |
Omar et al. | Conducting polymer and its composite materials based electrochemical sensor for Nicotinamide Adenine Dinucleotide (NADH) | |
Kaya et al. | Nanomaterials-based nanosensors for the simultaneous electrochemical determination of biologically important compounds: ascorbic acid, uric acid, and dopamine | |
Prasad et al. | Multi-wall carbon nanotube–NiO nanoparticle composite as enzyme-free electrochemical glucose sensor | |
Zhu et al. | Electrochemical detection of dopamine on a Ni/Al layered double hydroxide modified carbon ionic liquid electrode | |
Qu et al. | Magnetic loading of carbon nanotube/nano-Fe3O4 composite for electrochemical sensing | |
CN101982764B (en) | Composite membrane modified biosensor and preparation method and application thereof | |
Lin et al. | Performing enzyme-free H2O2 biosensor and simultaneous determination for AA, DA, and UA by MWCNT–PEDOT film | |
Ragupathy et al. | Electrocatalytic oxidation and determination of ascorbic acid in the presence of dopamine at multiwalled carbon nanotube–silica network–gold nanoparticles based nanohybrid modified electrode | |
Zhang et al. | Determination of dopamine in the presence of ascorbic acid by poly (styrene sulfonic acid) sodium salt/single-wall carbon nanotube film modified glassy carbon electrode | |
Xu et al. | Impedance DNA biosensor using electropolymerized polypyrrole/multiwalled carbon nanotubes modified electrode | |
Khan et al. | Electrochemical determination of uric acid in the presence of ascorbic acid on electrochemically reduced graphene oxide modified electrode | |
Ge et al. | Phosphorene nanocomposite with high environmental stability and antifouling capability for simultaneous sensing of clenbuterol and ractopamine | |
Wang et al. | Layer-by-layer assembly of copper nanoparticles and manganese dioxide-multiwalled carbon nanotubes film: a new nonenzymatic electrochemical sensor for glucose | |
CN104614425B (en) | A kind of functionalized carbon nano-tube carries Cu2The preparation and application of O hexagram crystallite composites | |
Erdem et al. | Single‐walled carbon nanotubes modified graphite electrodes for electrochemical monitoring of nucleic acids and biomolecular interactions | |
Xia et al. | A novel bio-electrochemical ascorbic acid sensor modified with Cu4 (OH) 6SO4 nanorods | |
CN103323516A (en) | Method for preparing non-enzyme H2O2 sensor based on ionic liquid functionalized carbon nanotube and silver nanometer compound | |
Wang et al. | Electrochemical parameters of ethamsylate at multi-walled carbon nanotube modified glassy carbon electrodes | |
Salimi et al. | Carbon Nanotubes‐Ionic Liquid and Chloropromazine Modified Electrode for Determination of NADH and Fabrication of Ethanol Biosensor | |
Chang et al. | Carboxymethyl cellulose assisted preparation of water-processable halloysite nanotubular composites with carboxyl-functionalized multi-carbon nanotubes for simultaneous voltammetric detection of uric acid, guanine and adenine in biological samples | |
CN103063715A (en) | Method for detecting surviving gene based on graphene-gold composite material electrochemical DNA (Deoxyribose Nucleic Acid) biosensor | |
CN103743805A (en) | Biosensor based on nitrogen-hybridized mesoporous carbon as well as preparation method and application of biosensor | |
Thangaraj et al. | A flow injection analysis coupled dual electrochemical detector for selective and simultaneous detection of guanine and adenine |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20130529 Termination date: 20170419 |
|
CF01 | Termination of patent right due to non-payment of annual fee |