CN102352413A - Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof - Google Patents
Nucleic acid nano-Au biosensor for detecting lead ions and preparation method thereof Download PDFInfo
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- CN102352413A CN102352413A CN2011103133730A CN201110313373A CN102352413A CN 102352413 A CN102352413 A CN 102352413A CN 2011103133730 A CN2011103133730 A CN 2011103133730A CN 201110313373 A CN201110313373 A CN 201110313373A CN 102352413 A CN102352413 A CN 102352413A
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Abstract
The invention discloses a nucleic acid nano-Au biosensor for detecting lead ions and a preparation method thereof. The nucleic acid nano-Au biosensor for detecting lead ions utilizes the principle that: when lead ions exist, the specific nuclease can cut the substrate which is complemented therewith, and utilizes the immune colloidal gold as an amplification color appearance system, and accordingly, the lead irons are quickly detected. The nucleic acid nano-Au biosensor for detecting lead ions has the advantages of high flexibility, low cost, no need of using any instrument and the like, not only the defect of requiring large-scale instruments in the former detection methods is solved, but also the detection flexibility can be ensured.
Description
Technical field
The present invention relates to the metal ion detection field, be specifically related to a kind of nucleic acid nano-gold biosensor that is used to detect lead ion and preparation method thereof.
Background technology
The method that detects lead ion at present both at home and abroad mainly contains spectroscopic analysis, electrochemical methods, mass spectrometry etc.
Spectroscopic analysis has comprised visible spectrophotometry, atomic absorption spectrometry, atomic fluorescence spectroscopy, atomic emission spectrometry etc.Visible spectrophotometry sensitivity is low, selectivity is not high, and needs to use highly toxic substance and have volatile organic solvent, and the experimenter is had the potential injury, has limited the application of this method.In the flame atomic absorption spectrometry with the experiment condition relative complex of graphite furnace atomic absorption spectrometry; Need use large-scale instrument, the former detection sensitivity is relatively low, though the latter has higher sensitivity; But receive the interference of other coexistent impurities easily, the accuracy that influence is measured.When atomic fluorescence spectroscopy utilizes lead ion and suitable reductive agent such as sodium borohydride to react, can form gaseous hydride, can carry out fluorimetric principle after the atomize and detect.Though this method all is being better than graphite furnace atomic absorption spectrometry aspect sensitivity and the anti-interference degree, but still needs the use of instrument.
Electrochemical method has reagent dosage few, manual dexterity reliably, fast, simple relatively, advantage such as be easy to popularize.Common electrochemical detection method has: polarography, anodic stripping voltammetry, potentiometric stripping etc.But the Traditional electrochemical method generally uses mercury film electrode or mercury solution to measure, and contaminate environment works the mischief to operator ' s health easily.Make this method also receive the restriction of application facet.Simultaneously, in recent years, utilize the specific enzyme of lead ion that screens also to emerge in an endless stream as the method that the basis is used for detecting, Zhu etc. have made up inhibition type lead ion electrochemical luminous sensor with ruthenium dipyridyl derivatives marker DNA zyme.Horses etc. are the molecular recognition material with DNAzyme, and lead ion is an analytic target, and the ruthenium dipyridyl derivatives is the electrochemiluminescence marker, have designed the biosensor that detects lead ion based on the catalytic electrochemiluminescence of " 8-17 " DNAzyme of lead ion.
Mass spectrometry: mass spectrum is a kind of common instrument that is used to detect, though that mass spectrograph has is highly sensitive, amount of samples is few, and analysis speed is fast, can realize the characteristics of multicomponent analysis, and instrument is relatively more expensive, uses and keep in repair relatively difficulty.Most of areas does not all have this instrument, aspect practical application, acquires a certain degree of difficulty.
Colloid gold particle is a kind of novel nano-material that occurs in recent years, is stabilized nano gold particle, because of its unique characteristic is applied to every field rapidly; Comprise being used for catalyzed reaction optical material, electron device; Biosensor, the cell imaging of pharmaceutical carrier and high-contrast etc.Research shows that the color of nano-Au solution is relevant with the size of the spacing of nm gold particles and nanometer gold aggregate.The spacing of nm gold particles is if obviously surpass its mean diameter, and nanometer gold is dispersion state, shows as redness on the macroscopic view; If this spacing is less than mean diameter, and then nanometer gold is prone to reunite, and is state of aggregation, shows as purple on the macroscopic view to blue.Utilize this character of nanometer gold, can design a series of biochemical reactions with the distance between the change nm gold particles, thereby realize detection the target material.
Summary of the invention
The objective of the invention is to the deficiency that exists according in the existing lead ion detection technique; A kind of nucleic acid nano-gold biosensor that can the rapid detection lead ion is provided; When utilizing lead ion to exist; Special nuclease can cut off and its this principle of complementary substrate; And utilize immune colloid gold as amplifying Color Appearance System, be used to detect the existence of lead ion.
Another purpose of the present invention is to provide the preparation method of above-mentioned nucleic acid nano-gold biosensor.
A further object of the invention is to provide the method for use of above-mentioned nucleic acid nano-gold biosensor.
Above-mentioned purpose of the present invention is achieved through following technical scheme:
Lead ion detects the principle of research:
2003, Brown etc. filtered out a kind of dependence Pb
2+Specific nuclease, called after 8-17 nuclease.In this research, we have chosen the original series and the substrate sequence of this nuclease.
At existing Pb
2+Rely on the basis of enzymic activity DNAzyme the synthetic following six sections sequences of concrete design:
First section is the specific nuclease sequence of lead ion, contains 36 bases (shown in SEQ ID NO:1);
Second section is and nuclease complementary substrate sequence, comprises 26 bases (shown in SEQ ID NO:2), and the 3' end is complementary fully with enzyme, and 5 ' end is longer slightly than enzyme sequence, and complementary sequence contains a deoxyribonucleotide A, and it is the key position of enzymic activity;
The 3rd section sequence is used for colloid gold label for the competition sequence, and sequence is (5 '-SH-SEQ ID NO:3-3 '); Its 5 ' end sulfydryl modification; Be marked on the colloid gold particle, its sequence is identical with second section substrate sequence, and nucleic acid detection line marked emulative with the substrate sequence combines;
The 4th section sequence drawn on detection line, and sequence is (5 '-biotin-SEQ ID NO:4-3 '), and the substrate of this section sequence and nuclease and competition sequence are complementary fully;
The 5th section and the 6th section is the Quality Control sequence.One is marked on the Radioactive colloidal gold; Sequence is (shown in 5 '-SH-SEQ ID NO:5), draws on nature controlling line for one, and sequence is (5 '-biotin-SEQ ID NO:6-3 '); Their mutual complementary pairing is used for the stability (Fig. 1) of detection of biological transmitter.
The lead ion detecting sensor comprises four parts, sample pad, gold mark pad, nitrocellulose filter and absorbent pad.Two lines are arranged on the nitrocellulose filter, be respectively detection line and nature controlling line.The Radioactive colloidal gold that is combined with the 3rd section and the 5th section sequence is sprayed on the gold mark pad, with Streptavidin with after being marked with the 4th section serial response of vitamin H, draws and on nitrocellulose filter, forms detection line; With Streptavidin with after being marked with the 5th section serial response of vitamin H, draw and on nitrocellulose filter, form nature controlling line.At first nuclease and substrate are mixed free substrate and the emulative nucleotide sequence that combines on the detection line of the nucleic acid of Radioactive colloidal gold marked, the binding site on the sealing detection line with 1:1.1.Testing sample is added wherein reaction.If contain lead ion in the testing sample; Then nuclease can cut off and its complementary substrate sequence; Free substrate sequence can reduce; Thereby reduce its closure; The nucleotide sequence of colloid gold particle marked and the nucleotide sequence on the detection line react; Colloid gold particle rests on the detection line, thereby makes detection line show red; Colloid gold particle continues to pass through nature controlling line forward, and the nucleotide sequence generation complementary pairing reaction on the 5th section nucleotide sequence of Radioactive colloidal gold marked and the nature controlling line makes nature controlling line show redness, positive result.If there is not lead ion in the testing sample; Then nuclease can not cut off its complementary substrate sequence; After the last appearance; Because the nucleic acid on the detection line is sealed by free substrate sequence fully; Then can not react with the sequence of colloid gold particle marked, detection line does not develop the color, but the sequence of colloid gold particle marked still can react with the sequence on the nature controlling line; It is red that nature controlling line shows, negative result.If nature controlling line does not develop the color, no matter whether detection line develops the color, illustrate that all problem has appearred in biosensor itself, the result is insincere, null result (Fig. 2).
Use this biosensor, can detect lead ion fast, work as Pb
2+Concentration is that detection line all shows macroscopic redness under the above condition of 500nM, and, along with Pb
2+The increase of concentration, the color of detection line are also deepened gradually.
The technological line of lead ion detection research as shown in Figure 3.
Compared with prior art, the present invention has following beneficial effect:
When the present invention utilizes lead ion to exist cleverly; Special nuclease can cut off and his this principle of complementary substrate; And utilize immune colloid gold as amplifying Color Appearance System, design a kind of highly sensitive, low expense, need not use the colloidal gold strip biosensor of any instrument.Not only on sensitivity and specificity, can reach requirement, and quick, easy, direct, significant to the detection of heavy metal ion.Both solved the defective that needs large-scale instrument in the detection method in the past, and can guarantee detection sensitivity again, and prepare easy, detect rapidly, do not need the technical professional.
Description of drawings
Fig. 1 is specific nuclease of lead ion and substrate thereof;
Fig. 2 is the synoptic diagram as a result after nucleic acid nano-gold biosensor according to the invention detects;
Fig. 3 is the experimental technique route map that nucleic acid nano-gold biosensor according to the invention detects lead ion;
Fig. 4 is the sensitivity figure as a result that nucleic acid nano-gold biosensor according to the invention detects lead ion;
Fig. 5 is the specificity figure as a result that nucleic acid nano-gold biosensor according to the invention detects lead ion.
Embodiment
Come further to explain the present invention below in conjunction with embodiment, but embodiment does not do any type of qualification to the present invention.
The preparation of embodiment 1 nucleic acid nano-gold biosensor according to the invention
1, the design of six kinds of nucleotide sequences
At existing Pb
2+Rely on the basis of enzymic activity DNAzyme, the synthetic six sections sequences of concrete design are shown in SEQ ID NO:1 ~ 6.
2. the preparation of nanometer gold (Radioactive colloidal gold):
In the round-bottomed flask of 250ml, take by weighing the HAuCL4 solution of 100g 0.01%, magnetic agitation is heated to boiling; In above-mentioned solution, add the trisodium citrate of 4ml 1% then rapidly, after solution becomes redness, continue to boil 10min, stop heating and continue to stir until cooling; Colloidal gold solution keeps in Dark Place for 4 ℃, and nanometer gold is identified through 520nm maximum absorbance value.
3. the preparation of gold mark nucleic acid:
With 100 μ l deionized water dissolving 1OD nucleotide sequences (get the 3rd and the 5th section nucleotide sequence, the mol ratio row of pressing 7:3 mix, and concentration is 1OD), join in 5 times of spissated colloidal gold solutions of volume of 1ml, 4 ℃ 24 hours; The sealing of 1% bovine serum albumin added the SDS of NaCl and 1% after 30 minutes, was that 0.15M and 0.01%, 4 ℃ spend the night to final concentration respectively, 11500 rev/mins centrifugal 20 minutes, abandon supernatant, sink to the bottom with the resuspended liquid of 500ul (20 mM Na
3PO
4, 5% BSA, 0.25% Tween and 10% sucrose) and resuspended, repeating to give a baby a bath on the third day after its birth suspends all over the resuspended liquid of back with 200ul again, processes suspension-s.
4. the processing of sample pad
Glass fibre soaks and contains pH9.0,4.0% TritonX-100,100mM boric acid, 3%PEG4000,1%BSA, 2% sucrose, and 0.1% SDS, 50.0nM competitor dna probe (SEQ ID NO:7), behind the solution of 0.5ug/ml salmon sperm dna, 37 ℃ of dry for standby.
5. the preparation of gold mark pad
The gold mark nucleotide sequence of the present invention preparation is applied on the glass fibre, and 37 ℃ of dryings 2 hours are processed gold mark pad, and are subsequent use.
6. the processing of nature controlling line and detection line on the nitrocellulose filter
Nucleotide sequence 6 with 32 μ l deionized water dissolving 1OD; Making its concentration is 100 μ M; Get the nucleotide sequence 6 of 15 μ l, 100 μ M; Add 15 μ l (1mg/ml) chain and sistomycocins; Reaction is after 2 hours under the room temperature; Adopt to draw a film metal spraying appearance and be applied on the nitrocellulose filter nature controlling line, 37 ℃ of dryings two hours.
Nucleotide sequence 4 with 32 μ l deionized water dissolving 1OD; Making its concentration is 100 μ M; Get the nucleotide sequence 4 of 15 μ l, 100 μ M; Add 15 μ l (1mg/ml) chain and sistomycocins; Reaction is after 2 hours under the room temperature; Adopt to draw a film metal spraying appearance and be applied on the nitrocellulose filter nature controlling line, 37 ℃ of dryings two hours.
7. the assembling of colloidal gold strip
Be fixed on the offset plate successively with being fixed with nitrocellulose filter, the thieving paper of oligonucleotide probe, the glass fibre that scribbles the nanoparticle labeled oligonucleotide probe, sample pad; The adjacent part 2mm that overlaps each other promptly obtains nucleic acid nano-gold biosensor of the present invention after cutting into wide 4mm.
Quality control standard (Fig. 2):
(1) it is effective that red line proof nano-gold biosensor appears in C line (nature controlling line).
(2) whether T line (detection line) red line occurs, is the positive negative standard of differentiating.
Criterion as a result:
(1) red line appears in the C line, and red line appears in the T line simultaneously, and Pb in the test sample is described
2+Content overproof;
(2) red line appears in the C line, and red line does not appear in the T line simultaneously, and Pb in the test sample is described
2+Content does not exceed standard;
(3) red line does not appear in the C line, illustrates that nano-gold biosensor lost efficacy.The quality control of nucleic acid nano-gold biosensor of the present invention and detection effect.
Embodiment 2
Adopt the made nucleic acid nano-gold biosensor of embodiment 1, carry out following experiment, verify that it detects effect.
1. prepare lead ion standardized solution gradient, concentration is respectively 100.0 μ M, 50 μ M, 10.0 μ M, 1.0 μ M, 500.0 nM, 100.0 nM, 0.0 nM, room temperature preservation.
2. prepare the Hg of 10.0 μ M
2+, Mn
2+, Cd
2+, Pb
2+, Mg
2+, Zn
2+, Fe
2+, Fe
3+, Ba
2+, Ca
2+, Ni2+, Co
2+Solution.
3. that gets 1 pipe, 1.0 OD respectively has specific nuclease of lead ion and a substrate, high speed centrifugation 10 seconds, and slowly uncap adds three respectively and boils off ionized water, and the concussion dissolving makes its final concentration be 100.0 μ M;
Nuclease and the substrate ratio row according to 1:1.1 are mixed, and with 4 * SSC dilution (pH8.0, NaCl replaces with NaNO3), the final concentration that makes nuclease is 1 μ M, and 70 ℃ of water-baths heated 2 minutes, placed warm water slowly to cool off, and this is a working fluid.
When detecting the Pb2+ in the aqueous solution, standard solution, the 8 μ L20 * SSC (pH8.0, NaCl replaces with NaNO3) with 6.0 μ L working fluids, 46.0 μ L different concns mix respectively, and the final concentration of nuclease and enzyme substrates is respectively 100.0 nM and 110.0 nM; , room temperature reaction 30 minutes.
4. that will dispose goes up the appearance drips of solution on sample pad, room temperature; Observe the colour-change of nature controlling line and detection line after 10 minutes.
The result shows: at Pb
2+Concentration is that detection line all shows macroscopic redness under the above condition of 500nM, and, along with Pb
2+The increase of concentration, the color of detection line are also deepened gradually, and be also consistent with the result of gold test strip bar readout instrument, along with Pb
2+The increase of concentration, the curve of detection line also increase gradually, the nature controlling line colour developing.At Pb
2+Under the condition that exists, its complementary substrate sequence of the specific incision of nuclease produces single-chain nucleic acid sequence to be detected, thus with the 3rd section sequence of Radioactive colloidal gold marked and detection line on the 4th section sequence of drawing form sandwich structure, make the apparent redness of detection line.And, Pb
2+Concentration high more, the efficient of nuclease is fast more, the substrate sequence of cut-out also increases thereupon, makes the color of detection line also deepen gradually.Simultaneously, nature controlling line also presents the redness of homogeneous, illustrates that the system of biosensor is normal, credible result.
5. respectively with the standard solution of 6 μ L working fluids and 46 μ L different ions, 8 μ L20 * SSC (pH8.0, NaCl replaces with NaNO3) mix, and make the final concentration of nuclease and enzyme substrates be respectively 100 nM and 110 n, room temperature reaction 30 minutes.
Above-mentioned reaction soln drops on the sample pad, room temperature; Observe the colour-change of nature controlling line and detection line after 10 minutes.
The result shows: under the condition of 10.0 μ M, have only Pb
2+Detection line show macroscopic redness, Ba
2+Faint redness is arranged, other Hg
2+, Mn
2+, Cd
2+, Mg
2+, Zn
2+, Fe
2+, Ca
2+, Ni
2+All do not develop the color.This result has demonstrated this biosensor has good specificity.Only at Pb
2+Under the condition that exists; Its complementary substrate sequence of the specific incision of nuclease ability; Thereby make the substrate sequence of sealing detection line reduce; Nucleotide sequence on the colloid gold particle and the sequence generation complementary pairing on the detection line; Macroscopic redness occurs, simultaneously, nature controlling line also presents the redness of homogeneous; The system that biosensor is described is normal, credible result (Fig. 4 ~ 5).
SEQ?ID?NO:1
CATCTCTTCTCCGAGCCGGTCGAAATAGTGAGTCAC
SEQ?ID?NO:2
CTCGTGACTCACTAT?AGGAAGAGATG
SEQ?ID?NO:3
TCACTATAG?GAAGAG
SEQ?ID?NO:4
CTCTTCCTATAGTGA
SEQ?ID?NO:5
ACAAAAAC?AAAAACA
SEQ?ID?NO:6
TGTTTTTGTTTT?TGT
SEQ?ID?NO:7
AGCTACGAGTT?GAGA
Claims (8)
1. a nucleic acid nano-gold biosensor that is used to detect lead ion is characterized in that comprising sample pad, gold mark pad, nitrocellulose filter and absorbent pad; Detection line and nature controlling line are arranged on the said nitrocellulose filter; Said detection line be with Streptavidin be marked with the SEQ ID NO:4 serial response of vitamin H after, draw and on nitrocellulose filter, to form; Said nature controlling line be Streptavidin be marked with the SEQ ID NO:5 serial response of vitamin H after, draw and on nitrocellulose filter, to form; Be sprayed with Radioactive colloidal gold on the said gold mark pad; Radioactive colloidal gold is combined with the sequence like SEQ ID NO:3 and SEQ ID NO:5.
2. the said preparation method who is used to detect the nucleic acid nano-gold biosensor of lead ion of claim 1; It is characterized in that being fixed in nitrocellulose filter, thieving paper, the glass fibre that scribbles the nanoparticle labeled oligonucleotide probe and sample pad on the offset plate successively; The adjacent part 2mm that overlaps each other is through cutting into the wide nucleic acid nano-gold biosensor of 4mm that is.
3. according to the said preparation method who is used to detect the nucleic acid nano-gold biosensor of lead ion of claim 2, the preparation method who it is characterized in that said colloidal gold solution is with 100g, 0.01% HAuCL
4Solution stirring is heated to boiling, adds 4ml, 1% trisodium citrate then, after solution becomes redness, continues to boil 10min, stops heating, continues to be stirred to cooling; After then the sequence of SEQ ID NO:3 and SEQ ID NO:5 being mixed according to certain molar ratio; Use deionized water dissolving, join in the 1-10 times of spissated colloidal gold solution of volume mixed mark 1-48h; Again with bovine serum albumin or the unnecessary avtive spot of PEG sealing; Add NaCl and SDS, respectively to final concentration be 0.1-0.15M and 0.01%-0.1%, aging after; Centrifugal 20 ~ the 60min of 8000 ~ 16000rpm; Abandon supernatant, resuspended deposition promptly obtains gold mark nucleic acid.
4. according to the said preparation method who is used to detect the nucleic acid nano-gold biosensor of lead ion of claim 2; It is characterized in that said sample pad is after the glass fibre immersion contains the mixing solutions of tensio-active agent, electric neutrality or ealkaline buffer (phosphoric acid buffer, borate buffer, Tris damping fluid, Hepes damping fluid etc.), PEG4000, BSA and sucrose, 37 ℃ of dry for standby.
5. according to the said preparation method who is used to detect the nucleic acid nano-gold biosensor of lead ion of claim 2, it is characterized in that said gold mark pad is gold to be marked nucleic acid be applied on the glass fibre, 37 ℃ of dry 30min ~ 3h process gold mark pad.
6. according to the said preparation method who is used to detect the nucleic acid nano-gold biosensor of lead ion of claim 2; It is characterized in that the nature controlling line on the said nitrocellulose filter is with deionized water dissolving nucleic acid sequence SEQ ID NO:6; Get the nucleic acid sequence SEQ ID NO:6 of certain volume again; Add chain and sistomycocin; Room temperature reaction 30min ~ 3h; Be applied on the nitrocellulose filter, 37 ℃ of dryings obtain nature controlling line.
7. according to the said preparation method who is used to detect the nucleic acid nano-gold biosensor of lead ion of claim 2; It is characterized in that the detection line on the said nitrocellulose filter is with deionized water dissolving nucleic acid sequence SEQ ID NO:4; Get the nucleic acid sequence SEQ ID NO:4 of certain volume again; Add chain and sistomycocin; Room temperature reaction 30min ~ 3h; Be applied on the nitrocellulose filter, 37 ℃ of dryings obtain detection line.
8. the said method of use that is used to detect the nucleic acid nano-gold biosensor of lead ion of claim 1; It is characterized in that said method is that nuclease (SEQ ID NO:1) and substrate (SEQ ID NO:2) are mixed with certain proportion; Free substrate and the emulative nucleotide sequence that combines on the detection line of the nucleic acid of Radioactive colloidal gold marked; Binding site on the sealing detection line adds wherein reaction with testing sample; If contain lead ion in the testing sample; Then nuclease can cut off and its complementary substrate sequence; Free substrate sequence can reduce; Thereby reduce its closure; The nucleotide sequence of colloid gold particle marked and the nucleotide sequence on the detection line react; Colloid gold particle rests on the detection line, thereby makes detection line show red; Colloid gold particle continues to pass through nature controlling line forward, and the nucleotide sequence generation complementary pairing reaction on the 5th section nucleotide sequence of Radioactive colloidal gold marked and the nature controlling line makes nature controlling line show redness, positive result; If there is not lead ion in the testing sample; Then nuclease can not cut off its complementary substrate sequence; After the last appearance; Because the nucleic acid on the detection line is sealed by free substrate sequence fully; Then can not react with the sequence of colloid gold particle marked, detection line does not develop the color, but the sequence of colloid gold particle marked still can react with the sequence on the nature controlling line; It is red that nature controlling line shows, negative result; If nature controlling line does not develop the color, no matter whether detection line develops the color, illustrate that all problem has appearred in biosensor itself, the result is insincere, is null result.
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| CN102643917A (en) * | 2012-04-25 | 2012-08-22 | 宁波大学 | Lead ion detection test paper and preparation method thereof |
| CN102735639A (en) * | 2012-06-29 | 2012-10-17 | 陕西师范大学 | Unlabeled-type homogeneous colorimetric method for detecting lead ions |
| CN103305605A (en) * | 2013-05-13 | 2013-09-18 | 中国科学院广州生物医药与健康研究院 | Non-enzyme ion detection method based on DNA (Deoxyribose Nucleic Acid) self assembly |
| CN105652012A (en) * | 2014-11-28 | 2016-06-08 | 惠州市银嘉环保科技有限公司 | Nanogold-DNA composite detection test paper for mercury ions, preparation method and use thereof |
| CN106546730A (en) * | 2016-10-28 | 2017-03-29 | 华中科技大学 | A kind of lead ion visible detection method |
| CN106636338A (en) * | 2016-10-19 | 2017-05-10 | 清华大学 | Method for detecting content of mercury ions and lead ions in to-be-detected solution at same time |
| CN106872690A (en) * | 2017-01-22 | 2017-06-20 | 武汉璟泓万方堂医药科技股份有限公司 | A kind of high sensitivity colloid gold test paper and preparation method and application |
| CN108251446A (en) * | 2018-01-12 | 2018-07-06 | 天津大学 | A kind of construction method of lead ion responsive type whole-cell biological sensor |
| CN114703256A (en) * | 2022-04-18 | 2022-07-05 | 中国农业科学院农业资源与农业区划研究所 | Detecting plant Pb2+DNAzyme fluorescent sensor |
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| CN102643917B (en) * | 2012-04-25 | 2013-11-27 | 宁波大学 | Lead ion detection test paper and preparation method thereof |
| CN102643917A (en) * | 2012-04-25 | 2012-08-22 | 宁波大学 | Lead ion detection test paper and preparation method thereof |
| CN102735639A (en) * | 2012-06-29 | 2012-10-17 | 陕西师范大学 | Unlabeled-type homogeneous colorimetric method for detecting lead ions |
| CN103305605A (en) * | 2013-05-13 | 2013-09-18 | 中国科学院广州生物医药与健康研究院 | Non-enzyme ion detection method based on DNA (Deoxyribose Nucleic Acid) self assembly |
| CN103305605B (en) * | 2013-05-13 | 2015-08-26 | 中国科学院广州生物医药与健康研究院 | Based on the non-enzymatic ion detection method of DNA self-assembly |
| CN105652012A (en) * | 2014-11-28 | 2016-06-08 | 惠州市银嘉环保科技有限公司 | Nanogold-DNA composite detection test paper for mercury ions, preparation method and use thereof |
| CN106636338B (en) * | 2016-10-19 | 2020-04-07 | 清华大学 | Method for simultaneously detecting content of mercury ions and lead ions in solution to be detected |
| CN106636338A (en) * | 2016-10-19 | 2017-05-10 | 清华大学 | Method for detecting content of mercury ions and lead ions in to-be-detected solution at same time |
| CN106546730A (en) * | 2016-10-28 | 2017-03-29 | 华中科技大学 | A kind of lead ion visible detection method |
| CN106546730B (en) * | 2016-10-28 | 2018-07-24 | 华中科技大学 | A kind of lead ion visible detection method |
| CN106872690B (en) * | 2017-01-22 | 2018-09-14 | 武汉璟泓万方堂医药科技股份有限公司 | A kind of high sensitivity colloid gold test paper and the preparation method and application thereof |
| CN106872690A (en) * | 2017-01-22 | 2017-06-20 | 武汉璟泓万方堂医药科技股份有限公司 | A kind of high sensitivity colloid gold test paper and preparation method and application |
| CN108251446A (en) * | 2018-01-12 | 2018-07-06 | 天津大学 | A kind of construction method of lead ion responsive type whole-cell biological sensor |
| CN114703256A (en) * | 2022-04-18 | 2022-07-05 | 中国农业科学院农业资源与农业区划研究所 | Detecting plant Pb2+DNAzyme fluorescent sensor |
| CN114703256B (en) * | 2022-04-18 | 2023-08-11 | 中国农业科学院农业资源与农业区划研究所 | Detection of plant Pb 2+ DNAzyme fluorescence sensor of (C) |
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