CN102375021A - Electrochemical method employing DNA as probe to detect environmental pollutant - Google Patents
Electrochemical method employing DNA as probe to detect environmental pollutant Download PDFInfo
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Abstract
An electrochemical method employing DNA as a probe to detect environmental pollutants. A biosensor detector comprises a working electrode, an auxiliary electrode, a reference electrode, a detection cell and an electrochemical working station. The working electrode is modified with nucleic acid probes on a surface thereof, and a genotoxicity compound in an environment sample is detected in a buffer system. When a detection result is positive, a signal of the sensor is changed obviously. Compared with an existing biosensor, the method has main advantages of: 1. high sensitivity; combination of an electrical active molecule and nucleic acid probes to increase a response current; 2. miniature and cheap apparatus and equipment, large detection flux, simple sample pre-treatment, and suitability for on spot rapid screening detection. The method can not only carry out screening detection on a known genotoxicity compound but also evaluate potential toxicity effect of a newly synthesized compound; and the method can screen whether an actual sample contains a genotoxicity compound.
Description
Technical field
The invention belongs to biochemical analysis and bio-sensing field, particularly relate to the method for determining presence of genotoxic compound in the DNA electrochemical sensor testing environment.
Background technology
The DNA sensor mainly is divided into the target DNA detection and dna mutation is measured two kinds.These are determined at, and aspects such as pathogen detection, food security, quarantine are widely used in human diseases diagnosis, the environment.
The growing interest that Along with people's is environmentally safe and healthy presses for and can carry out early diagnosis and therapy to various major diseases, rapidly surrounding enviroment is carried out safety evaluation.Yet, at present the examination that has the genotoxicity material in the environment is detected, mainly obtain the data a little less than its strong toxicity through animal experiment and cell toxicology evaluation, the method is costly, the cycle is long, is not suitable for the toxicity examination of compound quantitatively.Gas chromatography-mass spectrography (GC-MS) and liquid chromatograph mass spectrography (LC-MS); Can known toxic chemical qualitative, quantitative detect yet the large-scale costliness of its equipment is difficult to the scene of realizing, and test item be single; Flux is little, the sample pre-treatments complicated and time consumption.In addition, the method for chromatograph-mass spectrometer coupling can't be estimated unknown compound and the possible poisonous effect of noval chemical compound.
Because the shortcoming that animal experiment and chromatograph-mass spectrometer coupling experiment exist; After the seventies in 20th century; Chinese scholars begins to utilize microbiological sensor to detect dna damage or sequence change that the genotoxicity material causes; Mainly contain Ames method, SOS chromogenic reaction etc. and the method for therefrom deriving out thereof.Yet it is high that Ames method detects false positive; SOS chromogenic reaction insufficient sensitivity.
DNA electrochemical sensor used in the present invention has combined DNA and electrochemical advantage; On the whole; 1), specificity is good, has very high specific recognition capability through specific base pairing between the dna molecular two strands advantage that comprises the following aspects:.2), good stability, the DNA that exsomatizes is than the Heat stability is good of most protein (enzyme) molecule, the sensor of processing can be preserved the long period.3), preparation is simple, can synthesize in enormous quantities.4), electrochemical response is fast, operates easier.5), highly sensitive, with low cost, be fit to examination in enormous quantities and detect.
It is lower that yet electrochemical method directly detects the response signal that DNA produces, and faint response signal receives noise easily, and base-line shift takes place, and has reduced the sensitivity of instrument.And the concentration of determining presence of genotoxic compound is low in the environment, and the change in electric that causes is not obvious.
The sensitivity that the present invention uses the signaling molecule probe to increase the DNA electrochemical sensor, the change in electric that the feasible determining presence of genotoxic compound that is low to moderate picomole concentration produces also can be detected.
The invention provides a kind of simple and easy, fast and the screening technique of sensitive determining presence of genotoxic compound not only can be estimated a little less than the known toxic chemical strong toxicity; Can also estimate the poisonous effect of unknown compound; Whether contain determining presence of genotoxic compound in the examination actual sample.The instrument and equipment small inexpensive, it is big to detect flux, and sample pre-treatments is simple, is fit to on-site quick screening and detects.Can directly apply to the field screening and the detection of food and environmental sample, reach the purpose of economic, sensitive, accurate and examination fast in enormous quantities.
Summary of the invention
The purpose of this invention is to provide a kind of easy to operate, detection sensitivity is high, the result accurately and reliably, and can be used in the especially biology sensor detection method of determining presence of genotoxic compound of field screening testing environment pollutant.Be used for the DNA electrochemical sensor of determining presence of genotoxic compound examination, use it target analytes is detected, the result accurately and reliably, step is simple, and is highly sensitive, is fit to the field screening of batch samples.
Nucleic acid sensor detects the principle of determining presence of genotoxic compound: after the immobilized DNA effect on determining presence of genotoxic compound and the nucleic acid sensor; Through non-covalently combine with double-stranded DNA, mode such as DNA base damage can cause the structure of dna double chain or the slight change of pairing, above-mentioned variation can be carried out the signal amplification through on double-stranded DNA, introducing the electrochemical activity molecule methylene blue (MB) that combines with the DNA specificity.Can judge through the change in electric before and after record nucleic acid sensor and the determining presence of genotoxic compound effect whether target analytes has genotoxicity and toxicity size thereof.For single target analytes, the signal change intensity of this nucleic acid sensor and the concentration of target analytes is linear dependence within the specific limits.For complicated target sample such as water, soil, food, air, flue gas, whether this nucleic acid sensor can contain determining presence of genotoxic compound in the rapid screening target sample.This nucleic acid sensor can also be through the poisonous effect of or compound that toxicity unknown newly synthetic with the control experiment evaluation of toxicity compound known.
DNA capture probe and sealer (sulfydryl hexanol) that the present invention uses are fixed on gold electrode surfaces through end-labelled sulfydryl.The complementary target nucleic acid fragment 15～30 ℃ with gold electrode on immobilized DNA capture probe in buffer system (pH 7.0), reacted 1～2 hour.
The present invention adds the signaling molecule methylene blue (MB) with redox active that combines with the nucleic acid sensor characteristic in the detection solution of nucleic acid sensor, increased the response current signal of electrode.
The water-insoluble known compound that the present invention detects is dissolved in detection architecture and carries out the Electrochemical Detection analysis at cosolvent N under the effect of dinethylformamide.Unknown compound detects through electrochemical analysis, the genotoxicity effect of testing result comparative evaluation's unknown compound of its testing result and hexachloro-benzene.When target analytes is actual sample, can detect to confirm whether contain determining presence of genotoxic compound in the sample through electrochemical analysis.
Detection method of the present invention may further comprise the steps
A). an end of single stranded DNA capture probe is fixed on the working electrode surface of electrochemical appliance, obtains the trapping nucleic acids probe, adopt sealer to combine with the residue site of working electrode surface;
B). under hybridization conditions,, obtain the working electrode probe that double-stranded DNA is modified with complementary target dna fragmentation and above-mentioned trapping nucleic acids probe heterozygosis;
C). adopt the working electrode probe of the electroactive signaling molecule solution soaking double-stranded DNA modification that combines with the double-stranded DNA specificity, obtain nucleic acid sensor;
Nucleic acid sensor, contrast electrode and auxiliary electrode are placed the detection buffer system, carry out the electrochemical signals I that galvanochemistry scanning and record obtain
a, nitrogen dries up.Drip blank buffer solution on the modified electrode surface, handled 10～40 minutes, immerse the detection solution that contains 2～20 μ M signal probes (MB) subsequently, the scan-type electrochemical signal is made blank.；
D). take out nucleic acid sensor, the target analytes of liquid state is added drop-wise to the nucleic acid sensor surface, left standstill 10～30 minutes, buffer solution drip washing, the nucleic acid after obtaining handling is modified sensor;
Nucleic acid after will handling is then modified sensor and is immersed in above-mentioned detection buffer system again, carries out galvanochemistry scanning and recording responses signal I
b
E). through electric current decline number percent (I
a-I
b)/I
aWhether>=8%, determining presence of genotoxic compound is screened check and analysis; Electric current decline number percent through with n-compound (hexachloro-benzene) contrasts, and target analytes is carried out the poisonous effect evaluation;
Detect to confirm whether contain target analytes in the actual sample through the nucleic acid sensor electrochemical analysis; Described unknown compound is estimated the genotoxicity effect of unknown compound through the electric current decline number percent that nucleic acid sensor detects.
The preparation of nucleic acid sensor and the testing process of sample can be divided into following three steps:
The present invention uses peak current signal decline number percent assessing compound toxicity size: n is N, and dinethylformamide buffer solution is handled the peak current that scans behind the gold electrode of double-stranded DNA modification, i.e. the peak current of background solution scanning; M is 3,3 ', 4, the peak current that scans behind the gold electrode that detection compound solution-treated double-stranded DNAs such as 4 '-TePCB are modified; W is 3,3 ', 4, peak current signal decline number percent before and after the detection compound solution-treated such as 4 '-TePCB.Peak current signal decline number percent formula is following:
w＝[(n-m)/n]*100％
With respect to prior art, the present invention has following advantage:
1, testing result is reliable, accurate.The DNA sensor has better more stable to environment than protein (enzyme) sensor, thereby the compound that only has genotoxicity just can cause change in electric, has improved reliable, the accuracy of testing result.Can be used for whether containing determining presence of genotoxic compound in the examination actual sample.
2, enlarged sensing range.The use of non-genomic toxicity cosolvent makes a large amount of water-fast known organic contaminants list the sensing range of DNA sensor in; Can detect unknown solution simultaneously, determine whether to have genotoxicity.
3, can realize convenient, fast high-flux examination.Checkout equipment is made up of the electrochemical workstation of a microminiaturization, a three-electrode system and a detection cell, and equipment is simple, is easy to carry about with one, and is fit to the field condition examination.
Description of drawings
Fig. 1 is determining presence of genotoxic compound and a detector probe binding mode synoptic diagram in the inventive method.
Fig. 2 is monitoring of DNA probe and an electrode assembling process among the inventive method embodiment 1, and a is cyclic voltammetric (CV) curve of naked gold electrode in the 1mM potassium ferricyanide solution among the A figure, and b is the CV curve of DNA and sulfydryl hexanol modified gold electrode.A is for modifying the CV curve of gold electrode in 10 μ M MB solution of single stranded DNA (capture nucleic acid probe) among the B figure, and b is for modifying gold electrode CV curve in 10 μ M MB solution of double-stranded DNA.A gold electrode differentiated pulse scanning (DPV) curve in 10 μ M MB solution for modifying single stranded DNA in the illustration, b is for modifying gold electrode DPV curve in 10 μ M MB solution of double-stranded DNA.
Fig. 3 sweeps scanning curve under the speed for nucleic acid modified electrode difference in MB solution, is 80,60,40,20 from top to bottom, 10mV/s; Illustration is the linear correlation curve of sweep velocity and electric signal.
Fig. 4 is the curve of hexachloro-benzene concentration and the electric signal relation of the inventive method embodiment 2.
Fig. 5 is the UV, visible light curve of hexachloro-benzene and double-stranded nucleic acid probe effect among the inventive method embodiment 3.Be respectively: curve a representes MB solution; Curve b representes MB-DNA solution; Curve c representes 2 μ l 10
-2The hexachloro-benzene of M acts on MB-DNA.
Fig. 6 is the determining presence of genotoxic compound (100ng/ml) of the inventive method embodiment 4 and the electric signal decline number percent map that several kinds of non-genomic toxic chemicals (100ng/ml) cause: 1, potassium nitrate; 2, sodium citrate; 3, sodium oxalate; 4, urea; 5, ethyl acetate; 6, diethyl carbonate; 7, methenyl choloride; 8,3,3 ', 4,4 '-TePCB; 9,3,3 ', 4,4 ', 5,5 '-PePCB; 10,1,2,3,4,7,8-HxCDD; 11,2,3,7,8-TBrDF.
Embodiment
Following examples are used to explain the present invention, but are not used for limiting scope of the present invention.
Below be instrument and equipment used in the part embodiment of the invention, other not concrete experiment condition that indicates is according to the condition of routine or the suggestion of instrument manufacturing plant.
The used instrument of UV, visible light is ultraviolet-visible pectrophotometer (SP-1901, a China).The UV, visible light measuring condition: light source is ultraviolet and visible light, and light source automaticallyes switch at the 325nm place, and the scanning wavelength scope is 190～700nm, and sweep spacing is 1nm.Measure with the 3ml quartz colorimetric utensil, sample volume is 2ml, room temperature.The used instrument of Electrochemical Detection is Shanghai occasion China electrochemical workstation CHI440, three-electrode system, and detection architecture is in nitrogen atmosphere.Dna double chain (Mo Luonishi leukemia virus fragment) is all purchased in Takara (China, Dalian).Dna sequence dna is as shown in table 1.
Table 1 oligonucleotide base sequence
The assembling of embodiment 1. biological nucleic acid sensors
The gold electrode number of assembling steps:
1) surface area is 0.02cm
2Gold electrode is respectively 1,0.3 with particle diameter, the polishing of the alundum (Al powder of 0.05 μ m, ultrasonic cleaning 3 times repeatedly in absolute ethyl alcohol and deionized water then, each 2 minutes.Put into the scan round activation between 0～1.7V of 1M sulfuric acid solution, when scanning curve overlaps fully, stop scanning (activation completion).
2) cyclic voltammetry curve a representes among Fig. 2 A: the gold electrode of activation is put into the potassium ferricyanide buffer solution of 2mM, scan cycle volt-ampere curve between-0.1～0.6V.This curve redox spike potential difference explains that less than 75mV the potassium ferricyanide reaction of gold electrode surfaces belongs to the completely reversibility reaction, and gold electrode surfaces does not have impurity and quilt activation fully.After taking out electrode, dry up, drip the end modified single stranded DNA (trapping nucleic acids probe) of sulfydryl of 15 μ l, 2 μ M, room temperature treatment 6 hours at electrode surface with deionized water rinsing and nitrogen; Wash repeatedly with deionized water and 10mM PBS damping fluid (pH 7.0), nitrogen dries up, and adds 1mM sulfydryl hexanol (sealer) subsequently and handles 1 hour, residual activity site, enclosed-electrode surface.
3) CV curve b representes among Fig. 2 A: the gold electrode after trapping nucleic acids probe and sealer are modified is put into the potassium ferricyanide solution of 2mM, scan cycle volt-ampere curve between-0.1～0.6V.The peak current of the potassium ferricyanide greatly descends in the CV curve, explains that trapping nucleic acids probe and sealer have been attached to gold electrode surfaces fully.CV curve a representes among Fig. 2 B: the gold electrode of trapping nucleic acids probe modification immerses and contains in PBS (10mM) damping fluid of 10 μ M MB molecular probes, and nitrogen atmosphere stirred 10 minutes down, the interscan of-0.5～0V scope (sweep speed and be 100mV/s).CV curve b representes among Fig. 2 B: the gold electrode surfaces that the complementary chain dna of 15 μ l, 2 μ M is added drop-wise to the trapping nucleic acids probe modification is heterozygosis 2 hours at ambient temperature; Use deionized water and 10mM PBS damping fluid (pH 7.0) to wash repeatedly subsequently; After nitrogen dries up, in the PBS (10mM) of 10 μ M MB damping fluid, scan.Because the interaction of MB and strand capture probe is greater than interacting with double-stranded DNA, the binding capacity of MB and strand is greater than two strands, and the electric signal that the capture probe modified gold electrode produces is higher than the electric signal of double-stranded DNA modified gold electrode.
4) Fig. 3 for trapping nucleic acids probe modification gold electrode in 10 μ M MB solution with 10,20,40,60, five of 80mV/s different sweep speed scanning, draw that to sweep speed linear relevant with signal intensity, this reaction belongs to electrode surface and controls and react.
Embodiment 2. biological nucleic acid sensor hexachloro-benzenes
Use N, dinethylformamide is made cosolvent hexachloro-benzene is dissolved in the PBS damping fluid of 10mM, is made into the hexachloro-benzene solution of 100pM, 1nM, 10nM, 100nM, 1 μ M.
Fig. 4 representes: the gold electrode surfaces of modifying to double-stranded DNA drips 15 μ l hexachloro-benzene solution, and room temperature treatment 20 minutes immerses galvanochemistry scanning under the nitrogen atmosphere in PBS (10mM) damping fluid of 10 μ M MB then, obtains the correlation curve between concentration and the electric signal.Hexachloro-benzene and DNA effect, through non-covalently combine with double-stranded DNA, mode such as DNA base damage causes that the DNA pairing changes, and causes the nucleic acid sensor electric signal to change.Along with the rising of concentration, electric signal is progressively decayed, and signal change intensity and hexachloro-benzene concentration is linear dependence within the specific limits.
Fig. 5 representes: in quartz colorimetric utensil, add the PBS damping fluid of 2ml 10mM, full wavelength scanner is blank in 190～700nm scope, and curve a is the MB that in blank solution, adds 20 μ l 1mM, and full wavelength scanner obtains the MB peak.Curve b is the double-stranded DNA that in MB solution, adds 6 μ l, 100 μ M, shakes up and leaves standstill 10 minutes, and scanning obtains the peak that mixes of MB and DNA.The noiseless peak of selecting the 620-700nm place is for detecting wavelength coverage.Curve c adds 2 μ l 10 in the mixed solution of MB and DNA
-2The hexachloro-benzene of M is measured the absorption peak of solution at 620-700nm.
Obtained by interpretation: the MB peak appears at 190～350nm and 620～700nm, and the position at DNA peak is at 220～270nm, interferes with each other with the peak at MB 190～350nm place, and is unsuitable as the analysis and research object.And the peak of MB 620～700nm does not receive the peak interference of DNA, can be used as the analysis and research object of this experiment.After DNA added, MB embedded duplex DNA, and π-π takes place
*Conjugation, the MB peak intensity obviously weakens (hypochromic effect), and the position red shift 3nm of absorption peak.When adding 2 μ l 10
-2Behind the hexachloro-benzene of M, because strong interaction between hexachloro-benzene and DNA, hexachloro-benzene gets off part MB from the dna double chain substitution, and the absorption peak strength of MB increases (hyperchromicity), and blue shift occurs, when adding more than the 4 μ l 10
-2Behind the M hexachloro-benzene, blue shift and hyperchromicity no longer appear in the MB peak, and the combining of hexachloro-benzene and DNA reaches capacity.
Fig. 6 is for using N, and dinethylformamide is dissolved in the PBS of 10mM as cosolvent with following 12 kinds of compounds, is made into the solution of 100ng/ml, writes down change in electric with nucleic acid sensor; 12 kinds of detection compound are respectively: 1, potassium nitrate; 2, sodium citrate; 3, sodium oxalate; 4, urea; 5, ethyl acetate; 6, diethyl carbonate; 7, methenyl choloride; 8,3,3 ', 4,4 '-TePCB; 9,3,3 ', 4,4 ', 5,5 '-PePCB; 10,1,2,3,4,7,8-HxCDD; 11,2,3,7,8-TBrDF.Electrode surface in that double-stranded DNA is modified drips 15 μ l above-claimed cpd solution, and room temperature left standstill 20 minutes, immerses in PBS (10mM) damping fluid of 10 μ M MB scan-type electrochemical signal under the nitrogen atmosphere then.
Do not have the potassium nitrate, sodium citrate, sodium oxalate, urea, ethyl acetate, diethyl carbonate, methenyl choloride etc. of genotoxicity almost to detect and descend negative result less than electric signal.(obtain through data analysis: electric signal decline number percent was regarded as negative findings less than 8% o'clock).2,3,7,8-TBrDF and 1,2,3,4,7,8-HxCDD toxicity size is similar, and the electric signal decline number percent that the two causes is approaching.And PCB toxicity is less relatively, and the electric signal decline number percent that causes is less than 2,3,7,8-TBrDF and 1,2,3,4,7,8-HxCDD.Thereby reach a conclusion: varying in size of compound genotoxicity causes electric signal decline in various degree.
The method can significantly be distinguished background and have the response signal of the compound of genotoxicity, therefore can be used for doing the primary dcreening operation of environmental sample in enormous quantities; Not only can estimate, confirm the corresponding relation of its toxicity and electrical signal intensity, and can estimate the poisonous effect of unknown compound the examination of known toxic chemical; Whether contain determining presence of genotoxic compound in the examination actual sample.
Compare with the prior biological sensor, this method major advantage is: 1. highly sensitive, electroactive signaling molecule combines with nucleic acid probe, has increased response current; 2. instrument and equipment small inexpensive, it is big to detect flux, and sample pre-treatments is simple, is fit to on-site quick screening and detects.The method not only can be carried out examination to the known toxic chemical and detected, and can estimate the genotoxic potential effect of new synthetic compound; Whether contain determining presence of genotoxic compound in the examination actual sample.
Claims (9)
- One kind to adopt double-stranded DNA be the method for the Electrochemical Detection environmental contaminants of detector probe, adopt the electrochemical appliance of three-electrode system to detect, be characterised in that this method may further comprise the steps:A). an end of single stranded DNA capture probe is fixed on the working electrode surface of electrochemical appliance, obtains the trapping nucleic acids probe, adopt sealer to combine with the residue site of working electrode surface;B). under hybridization conditions,, obtain the working electrode probe that double-stranded DNA is modified with complementary target dna fragmentation and above-mentioned trapping nucleic acids probe heterozygosis;C). adopt the working electrode probe of the electroactive signaling molecule solution soaking double-stranded DNA modification that combines with the double-stranded DNA specificity, obtain nucleic acid sensor;Nucleic acid sensor, contrast electrode and auxiliary electrode are placed the detection buffer system, carry out the electrochemical signals I that galvanochemistry scanning and record obtain aD). take out nucleic acid sensor, the target analytes of liquid state is added drop-wise to the nucleic acid sensor surface, left standstill 10～30 minutes, buffer solution drip washing, the nucleic acid after obtaining handling is modified sensor;Nucleic acid after will handling is then modified sensor and is immersed in above-mentioned detection buffer system again, carries out galvanochemistry scanning and recording responses signal I bE). through electric current decline number percent (I a-I b)/I aWhether>=8%, determining presence of genotoxic compound is screened check and analysis; Electric current decline number percent through with n-compound (hexachloro-benzene) contrasts, and target analytes is carried out the poisonous effect evaluation;Detect to confirm whether contain target analytes in the actual sample through the nucleic acid sensor electrochemical analysis; Described unknown compound is estimated the genotoxicity effect of unknown compound through the electric current decline number percent that nucleic acid sensor detects.
- 2. the method for claim 1 is characterized in that: described double-strandednucleic acid is for containing 12～30 base-pairs, and the double-strandednucleic acid of guanine (G) content >=25%.
- 3. the method for claim 1, it is characterized in that: the working electrode of described electrochemical appliance is a noble metal electrode;Described trapping nucleic acids probe and sealer are fixed on electrode surface through end-labelled functional group; The functional group of described trapping nucleic acids probe and sealer is a kind of in biotin, siloxy, sulfydryl or the amino.
- 4. method as claimed in claim 3 is characterized in that: the working electrode of described electrochemical appliance is a gold electrode;The end-labelled functional group of described trapping nucleic acids probe and sealer is a sulfydryl.
- 5. method as claimed in claim 4 is characterized in that: said sealer is the end modified sealer of sulfydryl, i.e. sulfo-chain hydrocarbon or sulfo-aromatic hydrocarbon, preferably sulfo-chain hydrocarbon;Chain hydrocarbon in the described sulfo-chain hydrocarbon is meant that the carbon atom in the molecule combines the organic compound of chaining, and the aromatic hydrocarbon in the sulfo-aromatic hydrocarbon is meant the hydrocarbon that contains benzene ring structure in the molecule.
- 6. method as claimed in claim 5 is characterized in that: the carbon number number of described sulfo-chain hydrocarbon is 2～30, preferred 6～10 carbon atoms.
- 7. the method for claim 1 is characterized in that: described hybridization conditions is 0～60 ℃ of reaction 0.2～6 hour, preferred 15～30 ℃ of reactions 1～2 hour.
- 8. the method for claim 1 is characterized in that: the compound with redox active of described signaling molecule for combining with the DNA specificity, preferably methylene blue (MB) or bipyridyl ruthenium or dipyridine cobalt.
- 9. the method for claim 1, it is characterized in that: described target analytes is toxicity known compound or toxicity unknown compound;Described toxicity known compound is water soluble compound or the water-insoluble compound that dissolves in cosolvent; Water-insoluble compound is dissolved in buffer system under the effect of cosolvent; Described cosolvent is not for having the N of obvious influence, dinethylformamide to testing result;Described toxicity unknown compound is new synthetic compound or the unknown synthetic compound of poisonous effect.
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| CN104792978A (en) * | 2014-01-17 | 2015-07-22 | 中国科学院生态环境研究中心 | Signal labeled molecule for DNA oxidative damage product 8-hydroxydeoxyguanosine and labeling method |
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| CN110568036A (en) * | 2019-08-16 | 2019-12-13 | 成都理工大学 | Method for electrochemically detecting mercury ions based on nucleic acid dye |
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| CN116908265B (en) * | 2023-09-11 | 2023-12-12 | 常州先趋医疗科技有限公司 | Preparation method of electrochemical biosensor for detecting LAMP amplification products of nucleic acids |
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