CN106596676A - Electrochemical method for microRNAs detection - Google Patents
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Abstract
An electrochemical method for microRNAs detection comprises the following steps: A, nano silver solution preparation: B, working electrode preparation and C, electrochemical testing, the step B includes the steps of B1, electrodeposition of gold nanoparticles on the surface of a glassy carbon electrode; B2, assembling of a thiolated DNA capture probe 5'-TGACTTCAACATCAGTCTGATAAGCTAAGTCAT-(CH2) 6-SH-3' on the surface of the electrode prepared by the B1; B3, closing of the unreacted gold surface of the electrode prepared by the B2 with 6-mercapto-1-hexanol; B4, modification of the surface of the electrode prepared by the B3 with microRNAs; and B5, modification of the surface of the electrode prepared by the B4 with a nano silver solution mixture prepared by the step A and 4-mercaptophenylboronic acid, the step C electrochemical testing includes use of the electrode prepared by the B as a working electrode for electrochemical testing. The method has the advantages of simple operation, low cost, high sensitivity and accurate test.
Description
Technical field
The present invention relates to a kind of detection method of microRNAs, more particularly to a kind of electrification for microRNAs detections
Method, belongs to chemical field.
Background technology
Cancer is to threaten one of maximum killer of global human life, be that Medical research field is faced one
Significant challenge.The number of the infected of the annual cancer of China is about 2,000,000, dies from the number of cancer more than 1,400,000, and sickness rate is always
It is in rising trend.Cancer markers be reflect cancer chemical substance, early diagnosiss and treatment side of its detection in cancer
Mask is of great significance.Recently, research shows that the content of microRNAs in some Cancerous diseases there occurs substantially change
Change, therefore, microRNAs can be used as the biomarker of cancer.MicroRNAs is that raw, length is about 19- in a class
The single-stranded microRNA of 24 nucleotide.At present, the major technique for detecting miRNAs is blot hybridization technique, hybridization array skill
Art and PCR amplification techniques etc..The part however, these technologies come with some shortcomings, such as：Blot hybridization technique needs sample size
Greatly, sensitivity is low, and hybridization array technology needs expensive instrument, testing cost higher, and PCR amplification techniques are required experiment condition
Harsh, complex steps.Electrochemical sensor has that easy to use, certainty of measurement is high, maintenance is simple and low cost and other advantages, is based on
The property that the advantage of electrochemical techniques and nano material signal amplify, researcher was once attempted being realized using electrochemical means
The detection of microRNAs, but these methods need to carry out nano material surface modification, and complex steps, program are complicated, sensitivity
It is relatively low, it is unfavorable for the real-time detection of actual sample.
The content of the invention
It is an object of the invention to overcome the problems referred to above present in current microRNAs detections, there is provided one kind is used for
The electrochemical method of microRNAs detections.
To realize the purpose of the present invention, following technical schemes are employed：A kind of electrochemistry for microRNAs detections
Method, comprises the following steps：
A：The preparation of nanometer silver solution：Presoma is adopted for silver nitrate, reducing agent is sodium borohydride, stabilizer is sodium citrate,
Silver nitrate solution is added in sodium citrate solution, after being stirred vigorously 1 minute, sodium borohydride dissolving is added, is continued in room temperature
Under the conditions of stir 2 hours, that is, obtain the stable nanometer silver solution of citric acid, low temperature storage is standby after 2 days；
B：The preparation of working electrode, including following sub-step：
B1：In glassy carbon electrode surface electro-deposition gold nanoparticles；
B2：The DNA capture probes of sulfhydrylation are assembled in the electrode surface obtained by step B1：
5'-TGACTTCAACATCAGTCTGATAAGCTAAGTCAT-(CH2)6-SH-3'；
B3：The unreacted gold surface of electrode obtained by step B2 is closed using 6- sulfydryls hexanol；
B4：The electrode surface that microRNAs modifications are obtained to step B3；
B5：The electrode surface that the nanometer silver solution mixture modification that 4- mercaptophenyl boronic acids and step A are obtained is obtained to step B4,
Form nanometer silver network structure；
C：Electro-chemical test：The electrode that step B is prepared carries out electro-chemical test as working electrode；
Further, the nanometer silver solution cryopreservation that step A is obtained is standby, and storage temperature is 4 ~ 8 DEG C；
Further, a diameter of 3 mm of glass-carbon electrode in step B1；
Further, the concentration of the DNA capture probes in step B2 is 50 M, and the Tris bufferings using 100 mM Sodium Chloride are molten
Liquid is prepared, and the concentration of Tris is 20 mM, and pH value is 7.4；
Further, the ethanol of the 6- sulfydryls hexanol in step B3 dissolves, and 6- sulfydryl hexanols concentration is 1 mM；
Further, microRNAs adopts the Tris buffer preparations of 100 mM Sodium Chloride, Tris buffer solution in step B4
Concentration be 20 mM, pH value is 7.4, the redistilled water that described Tris buffer solution was processed using pyrocarbonic acid diethyl ester
Prepare；
Further, electrode obtained in step B4 is immersed in into the 4- mercaptophenyl boronic acid solution that concentration is 3.5 M in step B5
In, the nanometer silver solution that concentration is 1 nM is subsequently adding, distilled water flushing electrode is used in reaction after 30 minutes, remove unconjugated receiving
Rice silver particles；Described 4- mercaptophenyl boronic acids solution is prepared with PBS solution, and the concentration of PBS is 5 mM, and pH value is 7.4；
Further, electro-chemical test adopts three-electrode system in step C, and electrode prepared by step B is used as working electrode, saturation
Ag/AgCl electrodes be reference electrode, Pt electrodes be auxiliary electrode.
The Advantageous Effects of method provided by the present invention are：One of main distinction of RNA molecule and DNA molecular exists
In：Contain a hydroxyl on 2 ' numbers carbon of RNA molecule glycosyl, therefore, it is adjacent containing two in the glycosyl of its 3 ' terminal nucleotide
Cis hydroxyl groups,（I.e. cis vicinal diamines construction unit）, and vicinal diamines can form boric ester derivative with phenyl boronic acid derivative.
Additionally, citric acid is classical stabilizer the most frequently used when preparing gold silver nanoparticle, and citric acid also can be derivative with phenylboric acid
Thing forms the effect of boric acid ester bond.Based on these action characters, the present invention adopts 4- mercaptophenyl boronic acids（MPBA）As overseas Chinese federation's agent, knot
The electrochemical properties of nanometer silver are closed, a kind of simple, quick, electrochemical sensor of Sensitive Detection microRNAs is developed,
Contain vicinal diamines construction unit in the glycosyl of the 3 ' ends of microRNAs, can be with the boric acid base group phase in 4- mercaptophenyl boronic acids
Interaction, forms boric acid ester bond, it is achieved thereby that the specificity of microRNAs derives.Sulfydryl on 4- mercaptophenyl boronic acids can be with
The nanometer silver stable with citric acid forms Ag-S keys, and so as to nanometer silver is captured into electrode surface, the nanometer silver of electrode surface enters
One step is acted on by Ag-S keys and boric acid ester bond between phenylboric acid-citric acid is acted in adsorbent solution 4- mercaptophenyl boronic acids and
Nanometer silver, so as to more nanometer silvers are fixed to into electrode surface, so repeatedly, i.e., has been formed in situ nanometer silver in electrode surface
Network structure.The present invention captures microRNAs using DNA capture probes, is conducive to improving the specificity of analysis detection；Using 4-
Mercaptophenyl boronic acid derives to microRNAs, is conducive to acting on by Ag-S keys and the borate between phenylboric acid-citric acid
Multiple nano silver particles are fixed on electrode surface by key effect, and susceptiveness is high.The present invention without the need for being modified nano silver particles,
Nanometer silver network structure can be formed in electrode surface by way of growth in situ, simple to operate, cost is relatively low, detection time
Short, good stability.
Description of the drawings
Fig. 1 is the mass spectrum of the complex that 4- mercaptophenyl boronic acids are formed with citric acid reactions.
Fig. 2 is the mass spectrum of the complex that 4- mercaptophenyl boronic acids are formed with ribosidoadenine acid reaction.
Fig. 3 is the mass spectrum of the complex that 4- mercaptophenyl boronic acids are formed with cytidine acid reaction.
Fig. 4 is nanometer silver UV-visible absorption spectrum respectively in the case where 4- mercaptophenyl boronic acid existence conditions are whether there is.
Fig. 5 is the transmission electron microscope phenogram that 4- mercaptophenyl boronic acids induce nanometer silver aggregation.
Fig. 6 is electrode in process and the linear sweep voltammetry figure without the process of microRNA-21 modification steps.
Fig. 7 is linear sweep voltammetry figure when modifying the microRNA-21 of variable concentrations.
Fig. 8 is the graph of a relation of oxidation current and microRNA-21 concentration.
Fig. 9 is electrode in the linear sweep voltammetry figure through different microRNAs sequence modification steps.
Specific embodiment
In order to more fully explain the enforcement of the present invention, there is provided the embodiment of the present invention.These embodiments are only
Elaboration to the technique, does not limit the scope of the invention, and is illustrated with following examples in the present invention, but is not limited to following enforcements
Example, any change is included in the technical scope of the present invention.
Material in the present invention representated by various abbreviations is respectively：
5'-TGACTTCAACATCAGTCTGATAAGCTAAGTCAT-(CH2)6-SH-3'：DNA capture probes, 5'-
UAGCUUAUCAGACUGAUGUUGA-3'：MicroRNA-21, Tris：Trishydroxymethylaminomethane, PBS：Phosphate buffer,
MPBA：4- mercaptophenyl boronic acids, MCH：6- sulfydryl hexanols；MM, M, pM, fM are concentration unit, and 10 are represented respectively-3 mol/L、
10-6 mol/L、10-12 mol/L、10-15 mol/L.Following examples are carried out specifically using microRNA-21 as glycoprotein to be measured
Illustrate.DNA, microRNAs-21 for being related in application and the microRNA-21 of each base mispairing are biological by raw work
Engineering（Shanghai）Limited company's synthesis is provided.To accompanying drawing further explanation, in Fig. 4 curve a for nanometer silver it is ultraviolet-
Visible absorption spectra figure, curve b is UV-visible absorption spectrum of the nanometer silver under 4- mercaptophenyl boronic acid existence conditions.Fig. 6
In for electrode pass through（Curve a）With without（Curve b）The linear sweep voltammetry figure of microRNA-21 modification steps, curve a
The concentration of corresponding microRNA-21 is 2 pM, and supporting electrolyte is KCl.Corresponding microRNA-21 from top to bottom in Fig. 7
Concentration be followed successively by 0.1,1,10,20,50,100,200,500 and 2000 fM.Fig. 9 is that electrode is being passed through
microRNA-21（Curve a）, single base mismatch microRNA-21（Curve b）, three base mispairings microRNA-21（It is bent
Line c）, completely not complementary microRNA-21（Curve d）The linear sweep voltammetry figure of modification step, the concentration of microRNA-21
For 50 fM, single base mismatch, three base mispairings, the concentration of completely not complementary microRNA-21 are 500 fM.Single base
Mispairing, three base mispairings, the sequence of completely not complementary microRNA-21 are successively：5'-
UAGCUUAUCGGACUGAUGUUGA-3', 5'-UUGCUUAUCGGACUGAUCUUGA-3', 5'-
GUAAGGCAUCUGACCGAAGGCA-3'.Curve in Fig. 9 at peak valley is followed successively by from top to bottom curve d, curve c, curve b, song
Line a.
4- mercaptophenyl boronic acids and citric acid, adenylic acid, the case verification of the reaction of cytidylic acid：
By the 4- mercaptophenyl boronic acids solution of the M of 0.5 mL 200 citric acid, ribosidoadenine respectively with the M of 0.5 mL 200
Acid, cytidine acid solution mix 5 minutes, then flow injection carry out anion model measurement in mass detector.Figure
1 it is corresponding be 4- mercaptophenyl boronic acids-citric acid complex mass spectrum, illustration is the structural formula of the complex, 309.0238 Da
Corresponding charge-mass ratio when being one negative charge of composite belt.It is that 4- mercaptophenyl boronic acids-adenylic acid is multiple that Fig. 2 is corresponding
The mass spectrum of compound, illustration is the structural formula of the complex, and 463.0541 Da are corresponding when being one negative charge of composite belt
Charge-mass ratio.It is the mass spectrum of 4- mercaptophenyl boronic acids-cytidylic acid complex that Fig. 3 is corresponding, and illustration is the complex
Structural formula, the charge-mass ratio that 439.0417 Da are corresponding when being one negative charge of composite belt.These results show 4- sulfydryl benzene
Boric acid can react with citric acid, adenylic acid, cytidylic acid, form boric acid ester compound.
A kind of electrochemical method for microRNAs detections, comprises the following steps：
A：The preparation of the stable nanometer silver solution of citric acid：Volume ratio is adopted for 1:3 HNO3/ HCl mixed liquors justify 100 mL
Bottom flask, graduated cylinder are cleaned up, and are dried.Add 36.8 mL redistilled waters, the mM of 1 mL 10 toward round-bottomed flask successively
AgNO3With the mM sodium citrates of 1 mL 10, it is stirred vigorously 1 minute, adds the mM NaBH of 1.2 mL 104, continue to be stirred at room temperature
2 hours, finally the pale yellow solution is placed in 4 ~ 8 DEG C of refrigerator, it is standby after storing 2 days；
B：The preparation of working electrode：
B1：The preparation of golden nanometer particle modified electrode, will the glass-carbon electrode of a diameter of 3 mm be placed on the HAuCl4 of 1 mL 1%
In solution, electrode surface is finally rinsed in electro-deposition 60 seconds under the current potential of -0.25 V with redistilled water；
B2：The electrode surface obtained in B1 assembles the DNA capture probes of sulfhydrylation：
5'-TGACTTCAACATCAGTCTGATAAGCTAAGTCAT-(CH2)6- SH-3', for capturing microRNA-21, i.e.,
The electrode that B1 is obtained soaks 12 hours in the DNA capture probe solution of 50 M, then uses distilled water flushing electrode surface；It is dense
The DNA capture probes for 50 M are spent using the Tris buffer preparations comprising 100 mM Sodium Chloride, the concentration of Tris is 20
MM, pH value is 7.4, and wherein MCH ethanol is dissolved as the mM of concentration 1；
B3：Using 6- sulfydryl hexanols（MCH）Unreacted gold surface on the closing electrodes that obtain of B2, will step B2 obtain
Electrode soaks 30 minutes in the MCH solution of 1 mM, then with ethanol and distilled water flushing electrode surface, wherein 6- sulfydryls hexanol is used
Ethanol dissolves, and 6- sulfydryl hexanols concentration is 1 mM；
B4：The microRNA-21 solution that 50 L concentration are 2 pM is added drop-wise to into the electrode surface that B3 is obtained, is reacted 30 minutes,
Then distilled water flushing electrode surface is used, unreacted microRNA-21 is removed；MicroRNAs solution adopts 100 mM chlorinations
The Tris buffer preparations of sodium, the concentration of Tris buffer solution is 20 mM, and pH value is 7.4, and described Tris buffer solution is adopted
The redistilled water processed with pyrocarbonic acid diethyl ester is prepared；
B5：Electrode obtained in B4 is immersed in the 4- mercaptophenyl boronic acid solution that 20 L concentration are 3.5 M, 20 are subsequently adding
L concentration is the nanometer silver solution of 1 nM, and distilled water flushing electrode is used in reaction after 30 minutes, removes unconjugated nano silver particles；
4- mercaptophenyl boronic acids solution is prepared with PBS solution, and the concentration of PBS is 5 mM, and pH value is 7.4；Concentration is that the nanometer silver of 1 nM is molten
Liquid is prepared using PBS solution dilution, and the concentration of PBS is 5 mM, and pH value is 7.4；
C：The electrode that step B is obtained is placed in the KCl solution of 1 M, electro-chemical test is carried out using linear sweep voltammetry；
Electrochemical Detection adopts three-electrode system, and, used as working electrode, the Ag/AgCl electrodes of saturation are reference to electrode prepared by step B
Electrode, Pt electrodes are auxiliary electrode.
Test result as shown in fig. 6, curve a is cyclic voltammetry result of the working electrode through B1-B6 steps, in figure
Oxidation peak by nanometer galactic longitude electrochemical oxidation generate AgCl and produce.Curve b is working electrode through B1-B6 steps
Cyclic voltammetry result, but in B4 steps, the concentration of microRNA-21 is 0, and comparison curvess a and b can learn：The method
Can be used for the detection of microRNA-21.
Using the embodiment of different microRNA-21 concentration：
The concentration of microRNA-21 in step B4 is changed, the concentration that microRNA-21 is respectively adopted is 0.1,1,10,
20,50,100,200,500 and 2000 fM, the condition of other steps does not change, and test result is as shown in Figure 7.From figure
As can be seen that nanometer silver oxidation current increases with the increase of microRNA-21 concentration, the number of electrode adsorption nanometer silver is illustrated
Concentration of the amount depending on microRNA-21.Fig. 8 is the relation of oxidation current and microRNA-21 concentration.Can from figure
Go out, current intensity is with microRNA-21 concentration（0.1~50 fM）Increase and linearly increase, show that the method can be used for
The detection by quantitative of microRNA-21.Detection is limited to 0.02 fM.
Response implementation example to microRNA-21 similar sequences：
Change microRNA-21 in step B4 into other sequences to be tested, the condition of other steps does not change, and experimental result is such as
Shown in Fig. 9.It can be seen that the electrochemical signals that the microRNA-21 of single base mismatch is produced are far smaller than
The electric current that the electrochemical signals that microRNA-21 is produced, three base mispairings and completely unmatched microRNA-21 sequences are produced
It is negligible.Therefore, the method can be with selective enumeration method microRNA-21.Curve in figure is corresponding to be respectively
microRNA-21（Curve a）, single base mismatch microRNA-21（Curve b）, three base mispairings microRNA-21（It is bent
Line c）, completely not complementary microRNA-21（Curve d）.The concentration of microRNA-21 is 50 fM, and the concentration of other sequences is equal
For 500 fM.
In the present invention, the nanometer silver solution that 4- mercaptophenyl boronic acids are induced is carried out into aggregation test：By receiving prepared by step A
The PBS solution of the mM of meter Yin Yong 5 dilutes 4 times, then carries out ultraviolet-visible spectrum test.From fig. 4, it can be seen that synthesized receives
Meter Yin has a stronger absworption peak at 406 nm（Curve a）, after adding 4- mercaptophenyl boronic acids, the absworption peak at 406 nm
Intensity decreases, and a new absworption peak is occurred in that at 550 nm, corresponding is the absworption peak of nanometer silver aggregation.Fig. 5's
Transmission electron microscope picture spectrum shows that 4- mercaptophenyl boronic acids can couple together nanometer silver, forms nanometer silver network structure.
After embodiments of the present invention are described in detail, one of ordinary skilled in the art is clearly understood that, is not taking off
Various change and modification can be carried out under above-mentioned claim with spirit, all technical spirits according to the present invention are real to more than
Any simple modification, equivalent variations and modification that example is made are applied, the scope of technical solution of the present invention is belonged to, and the present invention is also not
It is limited to the embodiment of example in description.
SEQUENCE LISTING
<110>Anyang Teachers College
<120>A kind of electrochemical method for microRNAs detections
<130> 2010
<160> 5
<170> PatentIn version 3.3
<210> 1
<211> 33
<212> DNA
<213>Artificial sequence
<400> 1
tgacttcaac atcagtctga taagctaagt cat 33
<210> 2
<211> 22
<212> RNA
<213>Artificial sequence
<400> 2
uagcuuauca gacugauguu ga 22
<210> 3
<211> 22
<212> RNA
<213>Artificial sequence
<400> 3
uagcuuaucg gacugauguu ga 22
<210> 4
<211> 22
<212> RNA
<213>Artificial sequence
<400> 4
uugcuuaucg gacugaucuu ga 22
<210> 5
<211> 22
<212> RNA
<213>Artificial sequence
<400> 5
guaaggcauc ugaccgaagg ca 22
Claims (8)
1. a kind of electrochemical method for microRNAs detections, comprises the following steps：
A：The preparation of nanometer silver solution：Presoma is adopted for silver nitrate, reducing agent is sodium borohydride, stabilizer is sodium citrate,
Silver nitrate solution is added in sodium citrate solution, after being stirred vigorously 1 minute, sodium borohydride dissolving is added, is continued in room temperature
Under the conditions of stir 2 hours, that is, obtain the stable nanometer silver solution of citric acid, low temperature storage is standby after 2 days；
B：The preparation of working electrode, including following sub-step：
B1：In glassy carbon electrode surface electro-deposition gold nanoparticles；
B2：The DNA capture probes of sulfhydrylation are assembled in the electrode surface obtained by step B1：
5'-TGACTTCAACATCAGTCTGATAAGCTAAGTCAT-(CH2)6-SH-3'；
B3：The unreacted gold surface of electrode obtained by step B2 is closed using 6- sulfydryls hexanol；
B4：The electrode surface that microRNAs modifications are obtained to step B3；
B5：The electrode surface that the nanometer silver solution mixture modification that 4- mercaptophenyl boronic acids and step A are obtained is obtained to step B4,
Form nanometer silver network structure；
C：Electro-chemical test：The electrode that step B is prepared carries out electro-chemical test as working electrode.
2. it is according to claim 1 it is a kind of for microRNAs detection electrochemical method, it is characterised in that：Step A is obtained
The nanometer silver solution cryopreservation for arriving is standby, and storage temperature is 4 ~ 8 DEG C.
3. it is according to claim 1 it is a kind of for microRNAs detection electrochemical method, it is characterised in that：Step B1
In a diameter of 3 mm of glass-carbon electrode.
4. it is according to claim 1 it is a kind of for microRNAs detection electrochemical method, it is characterised in that：Step B2
In DNA capture probes concentration be 50 M, using the Tris buffer preparations of 100 mM Sodium Chloride, the concentration of Tris is
20 mM, pH value is 7.4.
5. it is according to claim 1 it is a kind of for microRNAs detection electrochemical method, it is characterised in that：Step B3
In 6- sulfydryls hexanol with ethanol dissolve, 6- sulfydryl hexanols concentration be 1 mM.
6. it is according to claim 1 it is a kind of for microRNAs detection electrochemical method, it is characterised in that：Step B4
Middle microRNAs adopts the Tris buffer preparations of 100 mM Sodium Chloride, and Tris buffer concentrations are 20 mM, and pH value is
7.4, described Tris buffer the redistilled water preparation that molten employing pyrocarbonic acid diethyl ester was processed.
7. it is according to claim 1 it is a kind of for microRNAs detection electrochemical method, it is characterised in that：Step B5
It is middle that electrode obtained in step B4 is immersed in the 4- mercaptophenyl boronic acid solution that concentration is 3.5 M, concentration is subsequently adding for 1 nM
Nanometer silver solution, reaction use distilled water flushing electrode after 30 minutes, the unconjugated nano silver particles of removing；Described 4- sulfydryls
Phenylboric acid solution is prepared with PBS solution, and the concentration of PBS solution is 5 mM, and pH value is 7.4.
8. it is according to claim 1 it is a kind of for microRNAs detection electrochemical method, it is characterised in that：In step C
Electro-chemical test adopts three-electrode system, and, used as working electrode, the Ag/AgCl electrodes of saturation are reference to electrode prepared by step B
Electrode, Pt electrodes are auxiliary electrode.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107543852A (en) * | 2017-08-29 | 2018-01-05 | 济南大学 | A kind of Electrochemiluminescsensor sensor based on functional metal organic framework materials |
| CN110117642A (en) * | 2018-02-07 | 2019-08-13 | 大连理工大学 | Specific recognition and detection method of the boric acid-nanoparticle system to RNA |
| WO2019153137A1 (en) * | 2018-02-07 | 2019-08-15 | 大连理工大学 | Specific recognition and detection method for ribonucleic acid using boronic acid modified nanoparticle system |
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| CN101986147A (en) * | 2010-10-18 | 2011-03-16 | 上海市七宝中学 | Application of electrochemical sensor |
| WO2013015650A2 (en) * | 2011-07-27 | 2013-01-31 | 부산대학교 산학협력단 | Cu-co alloy dendrite electrode and biosensor including same |
| CN104614417A (en) * | 2014-02-17 | 2015-05-13 | 安阳师范学院 | Electrochemical method for detecting glycoprotein |
| CN105738433A (en) * | 2016-01-28 | 2016-07-06 | 安徽师范大学 | Electrochemical transducer based on silver nanoparticles and preparing method and application thereof |
| CN106248771A (en) * | 2016-11-04 | 2016-12-21 | 北京农业信息技术研究中心 | The microelectrode biosensor of a kind of original position In vivo detection Mirnas of plant and application thereof |
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| CN107543852A (en) * | 2017-08-29 | 2018-01-05 | 济南大学 | A kind of Electrochemiluminescsensor sensor based on functional metal organic framework materials |
| CN110117642A (en) * | 2018-02-07 | 2019-08-13 | 大连理工大学 | Specific recognition and detection method of the boric acid-nanoparticle system to RNA |
| WO2019153137A1 (en) * | 2018-02-07 | 2019-08-15 | 大连理工大学 | Specific recognition and detection method for ribonucleic acid using boronic acid modified nanoparticle system |
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