CN104020198B - A kind of signal amplification technique electrochemical sensor detects the method for DNA - Google Patents
A kind of signal amplification technique electrochemical sensor detects the method for DNA Download PDFInfo
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Abstract
The present invention relates to a kind of signal amplification technique electrochemical sensor preparation method and application. First prepare gold-nano platinum particle, and be fixed in gold-nano platinum particle surface taking thionine as electrochemistry reagent, then by DNA probe by being fixed on nanoparticle surface, make electrochemical probe. Hairpin dna self assembly is obtained to electrochemical sensor in gold electrode surfaces simultaneously. Add target dna on electrochemical sensor time, target dna be assembled in the lip-deep hairpin dna part of gold electrode surfaces complementary pairing and open hairpin structure. Then an other hairpin dna chain replaces target chain DNA by Competition, thereby discharges target chain DNA. The object chain discharging continues to open other hairpin structure DNA of electrode surface, and so ringing has been realized recycling of target chain DNA, in conjunction with the electrochemical probe of preparation, and the mensuration of realization to target dna. Sensor of the present invention has very high selective and detection sensitivity; Gold-nano platinum particle is that carrier loaded thionine is the advantage that probe has electrochemical stability.<!--2-->
Description
Technical field
The invention belongs to analytical chemistry and technical field of electrochemistry, relate to a kind of signal amplification technique electrochemical sensor systemPreparation Method and application. In addition, the invention still further relates to the method that adopts described electrochemical sensor to detect DNA.
Background technology
Detect particular sequence DNA is very important in clinical diagnosis, gene therapy and other biological medical research.Therefore the method for, setting up the DNA of high sensitivity fast detecting particular sequence is very necessary.
Generally, detecting DNA all hybridizes to realize based on DNA. In order to read hybridization reaction, usual wayThat the part complementary series of target dna is carried out to mark. The general marks such as fluorescence, electrochemical luminescence, chemiluminescent substance that adoptThe part complementary series of target dna makes DNA probe, according to the signal strength measuring target dna of DNA probe after hybridization reactionConcentration. But, because the concentration of target dna in living things system is extremely low, must adopt suitable detection method to improve sensorSensitivity. Therefore, some DNA detection methods of amplifying based on signal are developed: enzyme is cut cycle signal amplifying technique [HunX.,LiuF.,MeiZ.H.,et.al.,Signalamplifiedstrategybasedontarget-inducedstrandreleasecouplingcleavageofnickingendonucleasefortheultrasensitivedetectionofochratoxinA,BiosensorsandBioelectronics,2013,39,145-151], roll ring amplifying technique [WenY.Q., XuY., MaoX.H., et.al., DNAzyme-BasedRolling-CircleAmplificationDNAMachineforUltrasensitiveAnalysisofMicroRNAinDrosophilaLarva, Anal.Chem.2012,84,7664-7669], polymerase chain reaction[HartmanM.R.,YangD.Y.,TranT.N.N.,et.al.,ThermostablebranchedDNAnanostructuresasmodularprimersforpolymerasechainreaction,AngewandteChemieInternationalEdition, 2013,52,8699 – 8702], ring mediated isothermal amplification [DhamaK.,KarthikK.,ChakrabortyS.,et.al.,Loop-mediatedisothermalamplificationofDNA(LAMP):Anewdiagnostictoollightstheworldofdiagnosisofanimalandhumanpathogens:Areview,PakistanJournalofBiologicalSciences,2014,17,151-166] etc. These methods respectively have its advantage, can meet in various degree the testing requirement to DNA, but sensitivity are not high. So mustBeard and hair exhibition is a kind of highly sensitive, simple, stable detection method.
Summary of the invention
In view of the deficiencies in the prior art, the object of the present invention is to provide a kind of signal amplification technique electrochemical sensor systemPreparation Method and application. And provide a kind of method that adopts described electrochemical sensor to detect DNA.
The object of the present invention is achieved like this:
The present invention realizes by following measures: a kind of signal amplification technique electrochemical sensor preparation method, itsFeature is to comprise the following steps:
(1) preparation of gold-nano platinum particle;
(2) preparation of electrochemical probe;
(3) preparation of electrochemical sensor;
The preparation of gold-nano platinum particle of the present invention comprises the following steps: used to preparation and reserve fund sol solutionGlass container (volumetric flask, brown wide-mouth bottle, round-bottomed flask) steep 30min with chloroazotic acid, then rinse dry for standby with intermediate water;In 1000mL round-bottomed flask, add the HAuCl of 50mL1mM4, be heated with stirring to boiling, then add fast 5mL38.8mMNatrium citricum (NaC6H5O7), when solution becomes claret, in solution, add successively 5mL4.0 × 10-3molL-1'sHPtCl6，15mL10gL-1PVP, being heated with stirring to boiling after continue heating 2h. Solution becomes sepia from claret, makesAuPtNPs (gold-nano platinum particle), is transferred to brown bottle, and 4 DEG C store for future use.
The preparation of electrochemical probe of the present invention comprises the following steps: first, get the centrifuge tube of 2mL, add 10 μ LDNA1 and 10 μ LpH8.2, the Tris-HCl cushioning liquid of 50mM, 10 μ LTCEP reaction 1h, in order to activate sulfydryl; Then, anotherGet 2mL centrifuge tube and add successively 1mL gold-nano platinum particle and 1000 μ L thionine solution reaction 0.5h, make thionine be adsorbed on gold-platinumOn nano particle; Gold-the nano platinum particle that has adsorbed thionine is joined in the sulfydryl DNA1 having activated, put into shaking table jog anti-Answer 16h. Sulfydryl DNA1 and gold-nano platinum particle are coupled together; Finally, under 15000 rotating speeds, centrifugal 30min, obtains rednessPrecipitation, and with 10mM, the phosphate buffer solution of pH8.0 washs three times, is dispersed in 10mM, in the phosphate buffer solution of pH8.0Obtain DNA1/Th/AuPtNPs electrochemical probe, 4 DEG C keep in Dark Place.
The preparation of electrochemical sensor of the present invention comprises the following steps:
1. the pretreatment of gold electrode
Gold electrode is through α-A1 of 0.05 μ m2O3After polishing powder polishing, rinse well with redistilled water, and at ultrasonic wave waterUltrasonic 5min in bath, dries up with high pure nitrogen; Employing three-electrode system detects, and working electrode is gold electrode, is platinum filament to electrodeElectrode, reference electrode is Ag/AgCl electrode, at K3[Fe(CN)6] in solution, it is 0～0.8V that voltage is set, and gold electrode is carried outCyclic voltammetric (CV) scanning; If redox current peak in 0～1000mV, has illustrated that electrode surface is processed good. NoAgain process, until meet the requirements; After having surveyed, rinse electrode with intermediate water, dry up electrode surface, for subsequent use.
2. golden nanometer particle modified gold electrode preparation
First get 10 μ LAuNPs (diameter 15nm) and be coated in gold electrode surfaces, obtain golden nanometer particle modified gold electrode(AuNPs/GE), being placed in lucifuge place dries naturally.
3. the pretreatment of hairpin dna
Get a certain amount of 10-6The DNA4 of M or sulfydryl DNA3, in 2mL centrifuge tube, are placed in 95 DEG C of constant temperature water baths and heat5min, is placed in immediately the cooling 1min of frozen water and obtains hairpin dna after taking-up.
4. the preparation of electrochemical sensor
Get 3 of the above-mentioned hairpin dnas of 10 μ L and be coated in AuNPs/GE surface, 4 DEG C of self assembly 24h; Then get 10 μ L10-4The mercapto of MBase ethanol drips in electrode surface, and then reaction 30min rinses twice with the phosphate buffer solution of 10mM, obtains electrochemical sensingDevice.
Utilize electrochemical sensor prepared by said method to detect a method for DNA content, comprise the steps:
(1) get again 10 μ L target dnas 2 and be added drop-wise to the electrode surface of electrochemical sensor, hatch after 2h at 37 DEG C, then get 10μ L hairpin dna 4 is added drop-wise to electrode surface. At 37 DEG C, hatch 4h, then use 10mM, the phosphate buffer solution of pH8.0 rinses twoInferior. Finally, get electrochemical probe prepared by 10 μ L and be added drop-wise to electrode surface, continue to hatch 1h.
(2) electrode system is inserted to 0.1M, in the phosphate buffer solution of pH7.4, taking above-mentioned gained gold electrode as work electricityThe utmost point, Ag/AgC1 (saturated KCl) electrode is reference electrode, platinum electrode is that electrode is surveyed to the signal of telecommunication (Ip/ A), carry out the survey of DNAFixed
Invention has been investigated electrode surface and has been dripped after hairpin dna 4, the impact of reaction time (Time/h) on current strength. AsShown in Fig. 2, first oxidation peak current increases along with the increase in reaction time, and in the time that the time reaches 4h, peak current no longer increases,And present the trend remaining unchanged. The peak optimization reaction time of selecting is 4h.
Also investigated electrochemical probe adsorption time impact on current strength on electrode, as shown in Figure 3, arrived 10In 60min, peak current increases along with the increase of adsorption time. In the time that adsorption time continues to increase, peak current substantially no longer becomesChange. Therefore, taking 60min as optimum adsorption time.
Remarkable result of the present invention
The present invention has studied the relation between variable concentrations DNA2 and electrochemical signals intensity, has obtained detecting the mark of DNA2Directrix curve, the range of linearity and linear equation.
Under optimum condition, the concentration of target dna 2 is 3.0 × 10-14～2.0×10-12Within the scope of M and current strengthBe certain linear relationship, its equation of linear regression is Ip＝6.6×105C+1.0×10-8(IpElectrochemical signals intensity; C isThe concentration of DNA2, n=9) (Fig. 4), and linearly dependent coefficient R=0.9991, detectability is 0.9 × 10-15M(3σ)。
With with 5 electrode pair concentration be 2.0 × 10-13M object DNA2 carries out respectively 17 evaluated errors and is respectively2.9%, 2.6%, 2.7%, 2.9%, 2.5%, in mensuration process, electrode surface is all without obscission. Taking golden nanometer particle asCarrier loaded thionine is that probe is to 2.0 × 10-13M object DNA2 carry out respectively 17 evaluated errors be respectively 9.3%,7.9%, 7.7%, 3.2%, 5.7%, in mensuration process, 4 electrode surfaces have obscission. Show with gold-platinum nanoparticleSon is that carrier loaded thionine is that probe has the advantages that stability is high.
Brief description of the drawings
Fig. 1. signal amplification technique is measured object schematic diagram.
Fig. 2. electrode surface drips after hairpin dna 4, the impact of reaction time on current strength.
Fig. 3. electrochemical probe is the impact of adsorption time on current strength on electrode.
Fig. 4. current strength and object concentration relationship figure.
Detailed description of the invention
Example below will illustrate method of operating of the present invention, but can not serve as limitation of the invention.
Example: signal amplification technique electrochemical sensor detects the method for DNA
1. experimental section
1.1 instruments and reagent
CHI660B electrochemical workstation (Shanghai Chen Hua instrument company); Anke-TGL-16C fly father-in-law's board supercentrifuge (onSea market An Ting scientific instrument factory); PHS-3D type acidometer (Shanghai Lei Ci instrument plant); Adopt three-electrode system: gold electrode and repairingGilding electrode is working electrode, and Ag/AgC1 (saturated KCl) electrode is reference electrode, and platinum electrode is to electrode.
Na2HPO4·12H2O, Tianjin red rock chemical reagent factory; Three (2-carboxyethyl) phosphonium salt hydrochlorate (TCEP, tris (2-Carboxyethyl) phosphinehydrochloride), gold chloride (HAuCl4), trisodium citrate (Na3C6H5O7) all purchaseIn Tianjin Bo Di Chemical Co., Ltd.; The potassium ferricyanide: K3[Fe(CN)6], Tianjin Bo Di Chemical Co., Ltd.; FerrocyanidePotassium: K4[Fe(CN)6]·3H2O, Tianjin Guang Cheng chemical reagent Co., Ltd; Alumina powder: α-A12O3, thionine (Th) purchased fromShanghai Aladdin reagent company; Polyvinylpyrrolidone (PVPK30) Tianjin Guang Cheng chemical reagent Co., Ltd.
DNA artificial sequence synthetic used (match Parkson, Beijing bioengineering Co., Ltd buys) is as follows.
Preferred DNA1 partial sequence is: 5`-TTTTTTATTCGATCCGGTGCTCTTATGCCC-HS-3`
Preferred DNA2 partial sequence is: 5 '-CAATAACTACCGGGCATTACTGGCCTT-3 '
Preferred DNA3 partial sequence is: 5 '-SH-C6-AAAGCCAGTAATGCCCGGTAGTTATTCCATCGTGTACAATAACTACCGGGCATAAGAGCACCCTTGTAC-3’
Preferred DNA4 partial sequence is: 5 '-TAGTTATTGTACACGATGGAATAACTACCGGGCATCCATCGTGTAC-3’。
1.2 experimental procedure
1.2.1 the preparation of gold-nano platinum particle
Preparation and reserve fund sol solution glass container (volumetric flask, brown wide-mouth bottle, round-bottomed flask) used are used to chloroazotic acidBubble 30min, then rinses dry for standby with intermediate water. In 1000mL round-bottomed flask, add the HAuCl of 50mL1mM4, stirBe heated to boiling, then add fast the natrium citricum (NaC of 5mL38.8mM6H5O7), when solution becomes claret, to solutionIn add successively 5mL4.0 × 10-3molL-1HPtCl6，15mL10gL-1PVP, being heated with stirring to boiling after continue heating2h. Solution becomes sepia from claret, makes AuPtNPs, is transferred to brown bottle, and 4 DEG C store for future use.
1.2.2 the preparation of electrochemical probe
First, get the centrifuge tube of 2mL, add 10 μ L10-5The DNA1 of M and 10 μ LpH8.2, the Tris-HCl of 50mM is slowDissolved liquid, the TCEP reaction 1h of 10 μ L10mM, in order to activate sulfydryl. Then, separately get 2mL centrifuge tube and add successively 1mLAuPtNPs and 1000 μ L10-4The thionine of M, solution reaction 0.5h, is adsorbed on gold-nano platinum particle thionine. To adsorb sulphurThe AuPtNPs of violet joins in the sulfydryl DNA1 having activated, and puts into shaking table jog reaction 16h. Make sulfydryl DNA1 and AuPtNPsCouple together. Finally, under 15000 rotating speeds, centrifugal 30min, obtains red precipitate, and with 10mM, the phosphoric acid buffer of pH8.0 is moltenLiquid washing three times, is dispersed in 10mM, obtains DNA1/Th/AuPtNPs electrochemical probe, 4 in the phosphate buffer solution of pH8.0DEG C keep in Dark Place.
1.2.3 the preparation of electrochemical sensor
The pretreatment of gold electrode
Gold electrode is through α-A1 of 0.05 μ m2O3After polishing powder polishing, rinse well with redistilled water, and at ultrasonic wave waterUltrasonic 5min in bath, dries up with high pure nitrogen. Employing three-electrode system detects, and working electrode is gold electrode, is platinum filament to electrodeElectrode, reference electrode is Ag/AgCl electrode, at K3[Fe(CN)6] in solution, it is 0～0.8V that voltage is set, and gold electrode is carried outCyclic voltammetric (CV) scanning. If redox current peak in 0～1000mV, has illustrated that electrode surface is processed good. NoAgain process, until meet the requirements. After having surveyed, rinse electrode with intermediate water, dry up electrode surface, for subsequent use. First getAuNPs prepared by 10 μ L drips and is coated in gold electrode surfaces, obtains golden nanometer particle modified gold electrode (AuNPs/GE), is placed in lucifuge place certainlySo dry.
The pretreatment of hairpin dna
Get a certain amount of 10-6The DNA4 of M or sulfydryl DNA3, in 2mL centrifuge tube, are placed in 95 DEG C of constant temperature water baths and heat5min, is placed in immediately the cooling 1min of frozen water and obtains hairpin dna after taking-up.
The preparation of electrochemical sensor
Get 3 of the above-mentioned hairpin dnas of 10 μ L and be coated in AuNPs/GE surface, 4 DEG C of self assembly 24h, then get 10 μ L10-4The mercapto of MThe sealing of base ethanol, the phosphate buffer solution with 10mM after electrode is dry rinses twice, obtains electrochemical sensor.
Electrochemical gaging target dna
Design the target dna 2 of one group of concentration gradient, be added drop-wise to respectively on electrochemical sensor, hatch after 2h at 37 DEG C, get10 μ L hairpin dnas 4 are added drop-wise to electrode surface, hatch 4h at 37 DEG C, then use 10mM, and the phosphate buffer solution of pH8.0 rinses twoInferior. Finally, get electrochemical probe prepared by 10 μ L and be added drop-wise to electrode surface, continue to hatch 1h. Then utilize cyclic voltammetry orDPV records current strength, according to target dna 2 concentration and the current strength relation calibration curve of mapping to obtain, and according to calibration curve meterCalculate equation of linear regression.
To be added on electrochemical sensor containing the sample drop of target dna 2, hatch after 2h at 37 DEG C, get 10 μ L hairpin dnas 4Be added drop-wise to electrode surface, hatch 4h at 37 DEG C, then use 10mM, the phosphate buffer solution of pH8.0 rinses twice. Finally, get 10Electrochemical probe prepared by μ L is added drop-wise to electrode surface, continues to hatch 1h. Then utilize cyclic voltammetry or DPV to record electric current strongDegree, according to current strength, target dna 2 content in recycling equation of linear regression calculation sample.
And adopting standard addition method to evaluate method, the sample determination rate of recovery is 97.0-106.0%, measures knotFruit is in table 1, and method of the present invention has the advantages that precision is high in target dna detects.
The present invention is the model electrochemical that the mechanism that recycles of based target DNA has been developed a kind of highly sensitive detection DNASensor. The sensor of development has following advantage: it is easy that gold-nano platinum particle is that carrier loaded thionine is that probe has overcome probeCome off and the unsettled defect of electrochemical signals; The circulation of target dna is amplified signal, the electrochemical sensing utensil of designThere are high stability, high sensitivity feature. Therefore, designed electrochemical sensor body in the research method that builds detection DNAReveal good development prospect.
Table 1. is containing target dna 2 sample analysis measurement results
| Numbering | Contenta,b | Standard sample addition | The amount of recording | The rate of recovery (%) |
| 1 | 7.6 | 5.0 | 12.5 | 98.0 |
| 2 | 9.2 | 10.0 | 19.8 | 106.0 |
| 3 | 12.4 | 10.0 | 22.7 | 103.0 |
| 4 | 15.6 | 15.0 | 30.2 | 97.0 |
| 5 | 23.9 | 25.0 | 50.6 | 102.7 |
| 6 | 49.0 | 50.0 | 98.2 | 98.4 |
a7 measurement results
bUnit: 10-14M
Claims (2)
1. signal amplification technique electrochemical sensor detects a method of DNA, comprises the following steps:
(1) electrochemical probe preparation method is characterized in that:
(a) get the centrifuge tube of 2mL, add the Tris-HCl cushioning liquid of the 50mM of 2～30 μ LDNA1 and 2～30 μ LpH8.2With 10 μ LTCEP reaction 1h, in order to activate sulfydryl; Then, separately get 2mL centrifuge tube add successively 1mL gold-nano platinum particle and100～2000 μ L thionine solution reaction 0.5h, are adsorbed on gold-nano platinum particle thionine;
(b) gold-nano platinum particle that has adsorbed thionine is joined in the sulfydryl DNA1 having activated, put into the reaction of shaking table jog16h, couples together sulfydryl DNA1 and gold-nano platinum particle; Finally, under 15000 rotating speeds, centrifugal 30min, obtains red sinkingForm sediment, and with 10mM, the phosphate buffer solution of pH8.0 washs three times, is dispersed in the phosphate buffer solution of the pH8.0 of 1mL10mMIn obtain DNA1/Th/AuPtNPs electrochemical probe;
(2) pretreatment of gold electrode: gold electrode is through 0.2 μ m, 0.03 μ m, α-A1 of 0.05 μ m2O3After polishing powder polishing, use secondaryDistilled water flushing is clean, and in ultrasound bath ultrasonic 5min, dry up for subsequent use with high pure nitrogen;
(3) golden nanometer particle modified gold electrode preparation: first get 2～30 μ LAuNPs solution and drip and be coated in gold electrode surfaces, obtain goldenNanoparticle Modified gold electrode AuNPs/GE, is placed in lucifuge place and naturally dries;
(4) pretreatment of hairpin dna: get a certain amount of 1.0 × 10-4M～1.0×10-7The DNA4 of M or sulfydryl DNA3 in 2mL fromIn core barrel, be placed in 95 DEG C of constant temperature water baths and heat 5min, after taking-up, be placed in immediately the cooling 1min of frozen water and obtain hair fastenerDNA；
(5) preparation of electrochemical sensor: get 3 of the above-mentioned pretreated hairpin dnas of 2～30 μ L and be coated in AuNPs/GE surface, 4DEG C self assembly 24h; Then get 10 μ L10-4The mercaptoethanol of M drips in electrode surface, reacts 30min, then uses the phosphoric acid of 10mMCushioning liquid rinses twice, obtains electrochemical sensor;
(6) drafting of calibration curve: get the target dna 2 of one group of concentration gradient, be added drop-wise to respectively on electrochemical sensor 37 DEG CUnder hatch after 2h, get 10 μ L hairpin dnas 4 and be added drop-wise to electrode surface, hatch 4h at 37 DEG C, then use 10mM, the phosphoric acid of pH8.0Cushioning liquid rinses twice; Finally, get electrochemical probe prepared by 10 μ L and be added drop-wise to electrode surface, continue to hatch 1h; Then profitRecord current strength with cyclic voltammetry or DPV, according to the relation of object concentration and the current strength calibration curve of mapping to obtain, andCalculate equation of linear regression according to calibration curve;
(7) sample determination: will be added on electrochemical sensor containing the sample drop of target dna 2, hatch after 2h at 37 DEG C, get 10 μ LHairpin dna 4 is added drop-wise to electrode surface, hatches 4h at 37 DEG C, then uses 10mM, and the phosphate buffer solution of pH8.0 rinses twice;Finally, get electrochemical probe prepared by 10 μ L and be added drop-wise to electrode surface, continue to hatch 1h; Then utilize cyclic voltammetry or DPVRecord current strength, according to current strength, target dna 2 content in recycling equation of linear regression calculation sample;
(8) partial sequence of described DNA1 is: 5`-TTTTTTATTCGATCCGGTGCTCTTATGCCC-HS-3`；
The partial sequence of described DNA2 is: 5 '-CAATAACTACCGGGCATTACTGGCCTT-3 ';
The partial sequence of described DNA3 is: 5 '-SH-C6-AAAGCCAGTAATGCCCGGTAGTTATTCCATCGTGTACAATAACTACCGGGCATAAGAGCACCCTTGTAC-3’；
The partial sequence of described DNA4 is: 5 '-TAGTTATTGTACACGATGGAATAACTACCGGGCATCCATCGTGTAC-3’。
2. the method for a kind of signal amplification technique electrochemical sensor detection DNA according to claim 1 contains at detection DNAApplication in amount.
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| CN104328192B (en) * | 2014-11-05 | 2021-03-30 | 南京大学 | Ribozyme amplified high-sensitivity electrochemical immunoassay method |
| CN104569101A (en) * | 2014-12-26 | 2015-04-29 | 北京科技大学 | DNA electrochemical biosensor and preparation method thereof |
| CN105675687B (en) * | 2016-03-23 | 2018-04-13 | 安徽师范大学 | A kind of preparation method of electrochemica biological sensor and the method for detecting dnmt rna activity |
| CN105784658B (en) * | 2016-03-29 | 2019-02-15 | 济南大学 | A kind of method of quick detection tumour correlation small peptide MUC1 |
| CN106248941B (en) * | 2016-07-14 | 2019-03-05 | 济南大学 | A kind of method of Sensitive Detection small peptide MUC1 |
| CN107121469B (en) * | 2017-04-28 | 2019-05-21 | 中南大学 | A kind of construction method of high stability nucleic acid sensor |
| CN108663354B (en) * | 2018-03-19 | 2021-05-14 | 安徽师范大学 | Electrogenerated chemiluminescence sensor constructed based on DNA-silver nanoclusters, and preparation and application thereof |
| CN108918628B (en) * | 2018-09-18 | 2020-11-10 | 安徽智时代科技有限公司 | Electrochemical sensor based on thionine-gold nanoparticle-aptamer compound for heavy metal ion detection |
| CN110412097B (en) * | 2019-08-12 | 2021-08-20 | 济南大学 | Preparation method of photoelectrochemical sensor for ultrasensitive detection of microRNA |
| CN110779970B (en) * | 2019-09-18 | 2022-04-12 | 南京农业大学 | Electrochemical detection method for chicken infectious bronchitis virus H120 strain |
| CN114231597A (en) * | 2021-11-29 | 2022-03-25 | 中国科学院苏州生物医学工程技术研究所 | Circulating tumor DNA detection method based on DNAzyme circulating activation |
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