CN106596484A - Method for detecting Hg<2+> - Google Patents
Method for detecting Hg<2+> Download PDFInfo
- Publication number
- CN106596484A CN106596484A CN201611127910.1A CN201611127910A CN106596484A CN 106596484 A CN106596484 A CN 106596484A CN 201611127910 A CN201611127910 A CN 201611127910A CN 106596484 A CN106596484 A CN 106596484A
- Authority
- CN
- China
- Prior art keywords
- detection
- biosensor
- rich
- detect
- stranded dna
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 30
- 239000010931 gold Substances 0.000 claims abstract description 25
- 229910052737 gold Inorganic materials 0.000 claims abstract description 25
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims abstract description 23
- 238000006243 chemical reaction Methods 0.000 claims abstract description 8
- 239000002539 nanocarrier Substances 0.000 claims abstract description 5
- 108020004414 DNA Proteins 0.000 claims description 24
- 102000053602 DNA Human genes 0.000 claims description 20
- 108020004682 Single-Stranded DNA Proteins 0.000 claims description 20
- 239000000463 material Substances 0.000 claims description 7
- 239000000523 sample Substances 0.000 claims description 7
- 239000006228 supernatant Substances 0.000 claims description 7
- 239000011324 bead Substances 0.000 claims description 6
- 230000010355 oscillation Effects 0.000 claims description 3
- 239000012488 sample solution Substances 0.000 claims description 3
- 238000000108 ultra-filtration Methods 0.000 claims description 2
- 238000001514 detection method Methods 0.000 abstract description 39
- 235000013305 food Nutrition 0.000 abstract description 9
- 230000036541 health Effects 0.000 abstract description 8
- 238000012544 monitoring process Methods 0.000 abstract description 5
- 230000009466 transformation Effects 0.000 abstract description 2
- 239000002091 nanocage Substances 0.000 abstract 1
- 239000000126 substance Substances 0.000 abstract 1
- 239000002585 base Substances 0.000 description 25
- 239000000243 solution Substances 0.000 description 11
- PYWVYCXTNDRMGF-UHFFFAOYSA-N rhodamine B Chemical group [Cl-].C=12C=CC(=[N+](CC)CC)C=C2OC2=CC(N(CC)CC)=CC=C2C=1C1=CC=CC=C1C(O)=O PYWVYCXTNDRMGF-UHFFFAOYSA-N 0.000 description 7
- 229940043267 rhodamine b Drugs 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 238000004587 chromatography analysis Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 230000007613 environmental effect Effects 0.000 description 4
- NJSSICCENMLTKO-HRCBOCMUSA-N [(1r,2s,4r,5r)-3-hydroxy-4-(4-methylphenyl)sulfonyloxy-6,8-dioxabicyclo[3.2.1]octan-2-yl] 4-methylbenzenesulfonate Chemical compound C1=CC(C)=CC=C1S(=O)(=O)O[C@H]1C(O)[C@@H](OS(=O)(=O)C=2C=CC(C)=CC=2)[C@@H]2OC[C@H]1O2 NJSSICCENMLTKO-HRCBOCMUSA-N 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 229910001385 heavy metal Inorganic materials 0.000 description 3
- QSHDDOUJBYECFT-UHFFFAOYSA-N mercury Chemical compound [Hg] QSHDDOUJBYECFT-UHFFFAOYSA-N 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 239000003513 alkali Substances 0.000 description 2
- 230000008512 biological response Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 230000009881 electrostatic interaction Effects 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 229910052753 mercury Inorganic materials 0.000 description 2
- 230000005012 migration Effects 0.000 description 2
- 238000013508 migration Methods 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- MPDGHEJMBKOTSU-YKLVYJNSSA-N 18beta-glycyrrhetic acid Chemical compound C([C@H]1C2=CC(=O)[C@H]34)[C@@](C)(C(O)=O)CC[C@]1(C)CC[C@@]2(C)[C@]4(C)CC[C@@H]1[C@]3(C)CC[C@H](O)C1(C)C MPDGHEJMBKOTSU-YKLVYJNSSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- JJWSNOOGIUMOEE-UHFFFAOYSA-N Monomethylmercury Chemical compound [Hg]C JJWSNOOGIUMOEE-UHFFFAOYSA-N 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000001479 atomic absorption spectroscopy Methods 0.000 description 1
- 238000001391 atomic fluorescence spectroscopy Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 231100000704 bioconcentration Toxicity 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000000356 contaminant Substances 0.000 description 1
- 238000013270 controlled release Methods 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 238000004993 emission spectroscopy Methods 0.000 description 1
- 238000000295 emission spectrum Methods 0.000 description 1
- 239000003344 environmental pollutant Substances 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000002795 fluorescence method Methods 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000001095 inductively coupled plasma mass spectrometry Methods 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 229920000554 ionomer Polymers 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000007885 magnetic separation Methods 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 231100000719 pollutant Toxicity 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000011895 specific detection Methods 0.000 description 1
- 238000009498 subcoating Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
- 238000003911 water pollution Methods 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Biochemistry (AREA)
- Pathology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention provides a method for detecting Hg<2+>. The method can be applied in high-sensitivity and high-selectivity Hg<2+> detection in the fields of environment monitoring, medical treatment and health, food safety, life science and the like. According to the method, Hg<2+> detection is conducted through a biosensor, the adopted biosensor is constructed by a nano-carrier, a fluorescent substance and single-chain DNA rich in T basic groups, the single-chain DNA rich in the T basic groups is assembled on the surface of a gold nanocage serving as the nano-carrier through the electrostatic action by serving as a Hg<2+> identification unit, and then the biosensor with a controllable releasing function is constructed. The single-chain DNA rich in the T basic groups generates configuration transformation due to a specific molecular identification reaction between Hg<2+> and the identification unit, then molecule doors are opened to release fluorescent molecules, and then fluorescent detection on Hg<2+> is achieved. Therefore, high-sensitivity and high-selectivity Hg<2+> detection can be achieved by means of the method.
Description
Technical field
The present invention relates to a kind of detection Hg2+Method, and in particular to one kind can in vivo, food, medicine and ring
Apply in border for detecting Hg2+Method.
Background technology
It is well known that heavy metal pollution causes increasingly serious impact to human body health and living environment.Hydrargyrum is
A kind of heavy metal with serious physiological-toxicity, with high animal migration and bioconcentration, is distributed widely in all kinds of surrounding mediums
In food chain, by migration and transformation, water body, soil, air and biosphere etc. are circulated in.Hg2+Wastewater Pollutant is
The important component part of water pollution is constituted, the electrodeless mercury contaminants entered in water can not be biodegradable, but participate in food
Chain is circulated, and can be the higher methyl mercury of toxicity by microorganism conversion, after being enriched with a large number in vivo through food chain (especially
Fish) human body is entered, the health of the mankind is seriously threaten, the infringement of central nervous system, hormonal system and brain can be caused,
Additionally, it may also can destroy human immune system, death is even resulted in.For this purpose, Environmental Protection Agency (EPA) and the European Community
The maximum permissible value for proposing Mercury in Drinking Water respectively is 10nM and 5nM.
Therefore, quick, sensitive, accurate Hg is set up2+Detection method not only facilitates environmental monitoring, simultaneously for clinical doctor
The numerous areas such as, life sciences, food hygiene, medicine and cosmetics also have important realistic meaning.
At present, Hg2+Detection method mainly include：Spectrophotography, Atomic Absorption/emission spectrometry, inductive etc.
Ionomer emission spectrum method (ICPMS), cold steam atomic fluorescence spectrometry, chromatography, electrochemical process etc., however, compared to high
Expensive instrument and reagent, numerous and diverse sample pretreatment, time-consuming testing process etc., fluorescence method is more suitable for Hg2+Detection, not only
Convenience, economic, quickly detection requirement are preferably met, meanwhile, also there is highly sensitive, high selectivity, in water ring
Border improvement, clinical medicine, creation conservation culture environment and guarantee people's health aspect are of great immediate significance.
In recent years, modern molecular biology technique and nanometer new material develop into Hg2+Highly sensitive, specific biological pass
Sense detection technique provides new thinking and advantage, emerges many new techniques, new method.As golden nanometer particle family
In the Cenozoic, gold nanometer cage once come out just caused extensive concern and research.Research shows, including Hg2+Interior many
Metal ion species can specifically with nucleotide " bridging ", be allowed to form stable metal ion-base pair.Wherein, Hg2+Energy
The DNA base of enough and mispairing interacts to T-T, forms stable T-Hg2+- T base pairs, this characteristic is in bio-sensing
Design shows huge potentiality with application aspect, is Hg2+Detection provide new, special, efficient detection platform.Using
The gold nanometer cage of positive surface charge modification is quiet with gold nanometer cage surface using the single stranded DNA rich in T bases as nano container
Electro ultrafiltration builds the detection Hg of biosensor2+Technology yet there are no document report.
The content of the invention
In order to overcome the shortcomings of that prior art is present, for the biology based on the single stranded DNA-gold nanometer cage rich in T bases
Sensor has no report, therefore, the first object of the present invention：Propose and build and a kind of new can be used to detect Hg2+Biology
Sensor, will gold nanometer cage and Hg2+Identification probe and positive surface charge modification technique combine, and have the biosensor
Can biological response molecule door, once add target molecule to be measured such as Hg2+, it will there is Hg2+With rich in T bases single stranded DNA it
Between specific molecular identification reaction, i.e., in T-Hg2+Under the coordination of-T, the single stranded DNA rich in T bases is transformed into similar sending out
The double-spiral structure of card-type, causes the single stranded DNA rich in T bases to depart from from gold nanometer cage surface, makes the molecule door in plugging cage hole
It is opened, and is released the guest molecule such as fluorescence molecule in hole, the fluorescence signal of system is strengthened；Therefore, may be used
By system fluorescence intensity increase indicating Hg2+Presence and its concentration；Using the biosensor, it is right to realize
Hg2+The detection of highly sensitive, high selectivity；The second object of the present invention：A kind of detection Hg is provided2+Biosensor preparation
Method；The third object of the present invention：There is provided a kind of using biosensor detection Hg2+Method.By inspection proposed by the present invention
Survey method is used for Hg2+Fluoroscopic examination, Hg can be significantly increased2+The sensitivity of detection and accuracy, are environmental monitoring, food
The application of the numerous areas such as product health, medicine and cosmetics provides new method with research.
The present invention is achieved through the following technical solutions goal of the invention.Detection Hg proposed by the present invention2+Bio-sensing
Device is, using gold nanometer cage as nano-carrier, using its hollow porous architectural characteristic, guest molecule to be loaded inside it for example glimmering
Optical molecule, in order to prevent leaking for fluorescence molecule, by single strand dna door of its surface-assembled rich in T bases, by Jenner
The hole closure on rice cage surface；Wherein, the described single stranded DNA rich in T bases is, through specially designed, to enable and Hg2+
There is the identification reaction of specificity, its base sequence is 3 '-TTT GTT TGT TGG CCC CCC TTC TTT CTT A-
5 ', it can be assembled into gold nanometer cage surface by electrostatic interaction, specifically can be by gold nanometer cage surface modification positive electricity
The method of lotus and be assembled into gold nanometer cage surface, preferably by positive charge dressing agent diallyl dimethyl ammoniumchloride
In the strata cation of gold nanometer cage surface modification one.
Preferably, above-mentioned detection Hg2+Biosensor, described fluorescence molecule is rhodamine B.
One kind prepares detection Hg proposed by the present invention2+Biosensor preparation method, comprise the steps：
(1) magnetic bead is mixed with gold nanometer cage solution, shaken at room temperature 8-12h, Magneto separate, removes supernatant, add sun from
Sub- coating material diallyl dimethyl ammoniumchloride solution, is shaken at room temperature overnight；
(2) Magneto separate, removes supernatant, adds fluorescent material rhodamine B solution, is shaken at room temperature overnight；
(3) the single stranded DNA solution rich in T bases is added, is shaken at room temperature overnight；
(4) Magneto separate, cleaning is obtained detection Hg2+Biosensor.
A kind of biosensor detection Hg using the present invention2+Method, step is as follows：
(1) by Hg2+Sample solution is added in the biosensor of the present invention, and 37 DEG C of constant temperature oscillations react 1-3h, in T-
Hg2+Under the coordination of-T, there is Hg2+With the specific molecular identification reaction of the single stranded DNA rich in T bases, make rich in T alkali
The single stranded DNA of base departs from from gold nanometer cage surface, and molecule door is opened, and the rhodamine B molecule in gold nanometer cage is released；
(2) the above-mentioned solution of Magneto separate, collects supernatant, detects its fluorescence signal.
Beneficial effects of the present invention：Detection Hg proposed by the present invention2+Biosensor be by gold nanometer cage with rich in T alkali
The single stranded DNA of base combines, and molecular biosciences door is formed by electrostatic interaction, using target molecule Hg2+It is special between molecule door
Property molecular recognition reaction, make the single strand dna door rich in T bases be opened, discharge fluorescence molecule, realize fluorescence letter
Number detection, the method makes Hg2+Detection sensitivity be significantly improved, be capable of achieving to target molecule Hg2+Highly sensitive, high selection
The detection of property.Detection Hg proposed by the present invention2+Biosensor there is simple structure, good stability, controllability is strong, fluorescence
The advantages of signal is sensitive, meanwhile, not by other common interference materials such as Pb2+, Zn2+, Cd2+, Ni2+, Fe3+, Ca2+, Cu2+Plasma
Impact, with high selectivity, can be applicable to the field Hg such as environmental monitoring, health care2+Fluoroscopic examination.Experimental result table
Detection Hg that is bright, being proposed using the present invention and prepared2+Biosensor can be 5.0 × 10-12～1.0 × 10-9It is real in the range of M
Now to Hg2+The detection of highly sensitive, high selectivity.Detection Hg2+Biosensor and preparation method thereof and detection technique in ring
The fields such as border monitoring, health care, food safety, life sciences have larger application potential and wide application prospect.
Description of the drawings
Fig. 1：Different Hg2+The fluorescence signal intensity of concentration.
Fig. 2：Hg2+The linear relationship of concentration and fluorescence signal intensity.
Specific embodiment
Specific embodiment according to the present invention is the following is, technical scheme is described further, but this
Bright protection domain is not limited to these embodiments.Every change or equivalent substitute without departing substantially from present inventive concept is included in this
Within the protection domain of invention.
The present invention is specifically described below by embodiment, but the present invention is not limited by following embodiments.
Experimental apparatus：THZ-82A gas bath constant temperature oscillators (medical apparatus and instruments factory of Jintan City)；F-4600 spectrofluorophotometers
(Hitachi, Japan)；Magnetic separation rack (thinks again happy chromatographic technique development centre in Tianjin).
Experiment reagent：3-4 μm of sulfydryl modification magnetic bead (thinking happy chromatographic technique development centre again in Tianjin)；Rhodamine B (I
Fourth)；Single stranded DNA rich in T bases：3 '-TTT GTT TGT TGG CCC CCC TTC TTT CTT A-5 ' (Shanghai life work lifes
Thing Engineering stock Co., Ltd), PBS buffer solution is 0.01M (pH 7.4, Na2HPO4-NaH2PO4)。
Embodiment 1：
One kind prepares detection Hg proposed by the present invention2+Biosensor method, comprise the steps：
(1) 20 μ L sulfydryl magnetic beads are taken, is uniformly mixed with 400 μ L gold nanometer cages after being cleaned with PBS buffer solution, shaken at room temperature
10h, Magneto separate is cleaned with PBS buffer solution, removes supernatant, is added in the complex of resulting magnetic bead-gold nanometer cage
200 μ L concentration for 11.664mg/mL diallyl dimethyl ammoniumchloride solution, shaken at room temperature 10h；
(2) the above-mentioned solution of Magneto separate, adds 100 μ L1.0 × 10 after being cleaned with PBS buffer solution-5Mol/L rhodamine Bs
PBS solution, shaken at room temperature 10h；
(3) it is 1.0 × 10 to add 10 μ L concentration to above-mentioned solution-5The single stranded DNA solution rich in T bases of M, 37 DEG C of vibrations
12h, Magneto separate removes supernatant, detects Hg2+Biosensor prepare complete；
Wherein, described sulfydryl magnetic bead is the commodity (thinking happy chromatographic technique development centre again in Tianjin) of purchase；Described
Gold nanometer cage press literature method obtain (G.D.Moon, S.W.Choi, X.Cai, W.Y.Li, E.C.Cho, U.Jeong,
L.V.Wang and Y.N.Xia.J.Am.Chem.Soc.2011,133,4762–4765)。
Embodiment 2:
One kind utilizes biosensor proposed by the present invention for Hg2+Detection, method is as follows：
(1) by Hg2+Sample solution is added to detection Hg proposed by the present invention2+Biosensor in, with PBS buffer it is molten
Liquid (PH=7.4) dilutes, 37 DEG C of constant temperature oscillation 2h, and Hg occurs2+Know with the specific molecular between the single stranded DNA rich in T bases
Do not react, i.e., in T-Hg2+Under the coordination of-T, the single stranded DNA rich in T bases is transformed into the Double helix knot of similar hair fastener type
Structure, causes the single stranded DNA rich in T bases to depart from from gold nanometer cage surface, and the molecule door for making plugging cage hole is opened, and makes in hole
Guest molecule rhodamine B be released；
(2) the above-mentioned solution of Magneto separate, collects supernatant, fluoroscopic examination, testing conditions：Excitation wavelength and launch wavelength are distinguished
For 530,573nm.
Fig. 1 is the Hg of variable concentrations2+Corresponding fluorescence signal intensity, Hg2+Concentration be respectively (0；1.0×10-13；5.0
×10-13；1.0×10-12；5.0×10-12；1.0×10-11；4.0×10-11；8.0×10-11；1.0×10-10；5.0×10-10；
1.0×10-9；1.0×10-8；1.0×10-7M)；Fig. 2 is Hg2+The linear relationship of concentration and fluorescence signal intensity.As a result show,
Hg2+Concentration is 5.0 × 10-12～1.0 × 10-9During M, fluorescence signal intensity and Hg2+The logarithm value of concentration is in good linear pass
It is that its linear equation is：FL=332.6791+64.6606lgCHg 2+(10-13M), linearly dependent coefficient is 0.9963.
The present invention in combination with molecular biotechnology, reacts nanotechnology by the molecular recognition of specificity, is capable of achieving
To Hg2+Highly sensitive, high selectivity detection.Specifically gold nanometer cage is repaiied with the single stranded DNA rich in T bases and positive surface charge
Decorations technology combines, make the biosensor have can biological response molecule door, once have Hg2+Exist, then Hg occurs2+With
Specific molecular identification reaction between single stranded DNA rich in T bases, i.e., in T-Hg2+Under the coordination of-T, rich in T bases
Single stranded DNA be transformed into the double-spiral structure of similar hair fastener type, cause the single stranded DNA rich in T bases de- from gold nanometer cage surface
From, the molecule door for making plugging cage hole is opened, and is released the guest molecule rhodamine B in hole, therefore, using the life
Thing sensor, can realize to Hg2+The detection of highly sensitive, high selectivity.
Biosensor with gold nanometer cage as carrier proposed by the present invention has simple structure, and good stability is controllable
Property it is strong, the advantages of fluorescence signal is sensitive, meanwhile, not by other common interference materials such as Pb2+, Zn2+, Cd2+, Ni2+, Fe3+, Ca2+,
Cu2+The impact of plasma, with high selectivity, can be used for Hg2+Highly sensitive, specific detection, be not only able to for environment prison
The detection of the field such as survey, health care, food safety, life sciences heavy metal provides new approaches and methods, meanwhile, based on this
The controlled release technologies of guest molecule are also cancer target in the nano-carrier of invention, drug delivery, the clinic of major disease examine
It is disconnected to carry out new thinking with treatment belt.
1
Sequence table
SEQUENCE LISTING
<110>Qingdao University of Science and Technology
<120>A kind of method of detection Hg2+
<160> 1
<210> 1
<211> 28
<212> DNA
<213>Artificial sequence
<400> 1
attctttctt ccccccggtt gtttgttt 28
Claims (5)
1. it is a kind of to detect Hg2+Method, it is characterised in that step is as follows：
(1) by Hg2+Sample solution is added to by magnetic bead, nano-carrier, fluorescent material and Hg2+The built-up biology of identification probe
In sensor, 37 DEG C of constant temperature oscillations react 1-3h；
(2) the above-mentioned solution of Magneto separate, collects supernatant, detects its fluorescence signal.
2. it is a kind of to detect Hg as claimed in claim 12+Method, it is characterised in that：Described biosensor be by magnetic bead,
Gold nanometer cage, fluorescent material and Hg2+Identification probe is built-up.
3. it is a kind of to detect Hg as claimed in claim 12+Method, it is characterised in that：Described Hg2+Identification probe is rich in T
The single stranded DNA of base, it and Hg2+The identification reaction of specificity can occur.
4. it is a kind of to detect Hg as claimed in claim 12+Method, it is characterised in that：Described Hg2+Identification probe is base sequence
It is classified as the single stranded DNA rich in T bases of 3 '-TTT GTT TGT TGG CCC CCC TTC TTT CTT A-5 '.
5. it is a kind of to detect Hg as claimed in claim 12+Method, it is characterised in that：Described Hg2+Identification probe is by quiet
Electro ultrafiltration is assembled into gold nanometer cage surface.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611127910.1A CN106596484B (en) | 2016-12-09 | 2016-12-09 | A kind of detection Hg2+Method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201611127910.1A CN106596484B (en) | 2016-12-09 | 2016-12-09 | A kind of detection Hg2+Method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106596484A true CN106596484A (en) | 2017-04-26 |
| CN106596484B CN106596484B (en) | 2019-06-25 |
Family
ID=58598260
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201611127910.1A Active CN106596484B (en) | 2016-12-09 | 2016-12-09 | A kind of detection Hg2+Method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106596484B (en) |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106990081A (en) * | 2017-04-10 | 2017-07-28 | 江苏大学 | One kind is based on graphene oxide sensor and its to Hg2+The method of detection |
| CN110455897A (en) * | 2019-08-29 | 2019-11-15 | 济南大学 | One kind being based on SiO2Carrier Sensitive Detection Hg2+Release type electrochemical aptamer sensor building |
| CN110819695A (en) * | 2018-08-13 | 2020-02-21 | 青岛科技大学 | Method for detecting silver ions |
| CN111778244A (en) * | 2020-05-29 | 2020-10-16 | 南京邮电大学 | DNA probe combined with gold nanosphere and application of DNA probe in detection of Hg based on solid-state nanopore2+In (1) |
| CN113252622A (en) * | 2020-02-12 | 2021-08-13 | 青岛科技大学 | Hg simultaneous detection based on single excitation2+And Ag+Method (2) |
| CN113252620A (en) * | 2020-02-12 | 2021-08-13 | 青岛科技大学 | Hg can be detected simultaneously2+And Ag+Method (2) |
| CN113252619A (en) * | 2020-02-12 | 2021-08-13 | 青岛科技大学 | Hg can be detected simultaneously2+And Ag+The nanocapsule-nucleic acid biomolecule compound and the preparation method thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108593558B (en) * | 2018-04-04 | 2020-12-01 | 深圳大学 | For Hg2+Detected photoacoustic detection probe and preparation method thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101424642A (en) * | 2008-11-14 | 2009-05-06 | 中国科学院上海应用物理研究所 | Target molecule detecting method based on nanometer aurum and nucleic acid structure |
| CN101887020A (en) * | 2010-07-09 | 2010-11-17 | 南京邮电大学 | Fluorescence detection method of mercury ions by oligonucleotide and graphene oxide |
| CN101892294A (en) * | 2010-01-22 | 2010-11-24 | 华南师范大学 | Magnetic bead electrochemiluminescence gene sensor-based method for detecting mercury ions and application thereof |
| CN102200510A (en) * | 2011-04-13 | 2011-09-28 | 上海出入境检验检疫局机电产品检测技术中心 | Mercuric ion concentration fluorescence detection method based on T-T mismatched DNA probe |
| US8062893B2 (en) * | 2008-10-10 | 2011-11-22 | The Board Of Trustees Of The University Of Illinois | Fluorescent sensor for mercury |
| KR20120052918A (en) * | 2012-04-23 | 2012-05-24 | 한국과학기술원 | Method for detecting metal ions using unnatural polymerase activity and logic gate using the same |
-
2016
- 2016-12-09 CN CN201611127910.1A patent/CN106596484B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8062893B2 (en) * | 2008-10-10 | 2011-11-22 | The Board Of Trustees Of The University Of Illinois | Fluorescent sensor for mercury |
| CN101424642A (en) * | 2008-11-14 | 2009-05-06 | 中国科学院上海应用物理研究所 | Target molecule detecting method based on nanometer aurum and nucleic acid structure |
| CN101892294A (en) * | 2010-01-22 | 2010-11-24 | 华南师范大学 | Magnetic bead electrochemiluminescence gene sensor-based method for detecting mercury ions and application thereof |
| CN101887020A (en) * | 2010-07-09 | 2010-11-17 | 南京邮电大学 | Fluorescence detection method of mercury ions by oligonucleotide and graphene oxide |
| CN102200510A (en) * | 2011-04-13 | 2011-09-28 | 上海出入境检验检疫局机电产品检测技术中心 | Mercuric ion concentration fluorescence detection method based on T-T mismatched DNA probe |
| KR20120052918A (en) * | 2012-04-23 | 2012-05-24 | 한국과학기술원 | Method for detecting metal ions using unnatural polymerase activity and logic gate using the same |
Non-Patent Citations (2)
| Title |
|---|
| PAUL D. SELID等: "Sensing Mercury for Biomedical and Environmental Monitoring", 《SENSORS》 * |
| 宇佳 等: "基于核酸适配体与金纳米粒子检测水中Hg2+的研究", 《分析检测》 * |
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106990081A (en) * | 2017-04-10 | 2017-07-28 | 江苏大学 | One kind is based on graphene oxide sensor and its to Hg2+The method of detection |
| CN110819695A (en) * | 2018-08-13 | 2020-02-21 | 青岛科技大学 | Method for detecting silver ions |
| CN110819695B (en) * | 2018-08-13 | 2022-11-08 | 青岛科技大学 | Method for detecting silver ions |
| CN110455897A (en) * | 2019-08-29 | 2019-11-15 | 济南大学 | One kind being based on SiO2Carrier Sensitive Detection Hg2+Release type electrochemical aptamer sensor building |
| CN110455897B (en) * | 2019-08-29 | 2021-07-02 | 济南大学 | Based on SiO2Sensitive detection of Hg by carrier2+Construction of the Release electrochemical aptamer sensor |
| CN113252622A (en) * | 2020-02-12 | 2021-08-13 | 青岛科技大学 | Hg simultaneous detection based on single excitation2+And Ag+Method (2) |
| CN113252620A (en) * | 2020-02-12 | 2021-08-13 | 青岛科技大学 | Hg can be detected simultaneously2+And Ag+Method (2) |
| CN113252619A (en) * | 2020-02-12 | 2021-08-13 | 青岛科技大学 | Hg can be detected simultaneously2+And Ag+The nanocapsule-nucleic acid biomolecule compound and the preparation method thereof |
| CN111778244A (en) * | 2020-05-29 | 2020-10-16 | 南京邮电大学 | DNA probe combined with gold nanosphere and application of DNA probe in detection of Hg based on solid-state nanopore2+In (1) |
| CN111778244B (en) * | 2020-05-29 | 2023-10-27 | 南京邮电大学 | DNA probe combined with gold nanospheres and detection of Hg based on solid-state nanopores 2+ Application in (a) |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106596484B (en) | 2019-06-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106596484A (en) | Method for detecting Hg<2+> | |
| CN106770107B (en) | A kind of detection Hg2+Biosensor and preparation method thereof | |
| CN107064080B (en) | A kind of intracellular Hg2+Fluorescence imaging method | |
| Khoshbin et al. | Aptasensors as the future of antibiotics test kits-a case study of the aptamer application in the chloramphenicol detection | |
| Zhou et al. | SERS detection of bacteria in water by in situ coating with Ag nanoparticles | |
| Yang et al. | Microwave-assisted synthesis of polyamine-functionalized carbon dots from xylan and their use for the detection of tannic acid | |
| Liu et al. | SERS substrate fabrication for biochemical sensing: Towards point-of-care diagnostics | |
| Baghbaderani et al. | Novel chitosan-Nafion composite for fabrication of highly sensitive impedimetric and colorimetric As (III) aptasensor | |
| Huang et al. | Black phosphorus-Au filter paper-based three-dimensional SERS substrate for rapid detection of foodborne bacteria | |
| Mishra et al. | On-chip micro-biosensor for the detection of human CD4+ cells based on AC impedance and optical analysis | |
| CN104931684B (en) | Nano fluorescence sensor and preparation method and application thereof | |
| Chow et al. | Design and synthesis of heterobimetallic Ru (II)–Ln (III) complexes as chemodosimetric ensembles for the detection of biogenic amine odorants | |
| Wei et al. | Synthesis of molecularly imprinted silica nanospheres embedded mercaptosuccinic acid-coated CdTe quantum dots for selective recognition of λ-cyhalothrin | |
| CN106908429B (en) | Method for detecting glutathione | |
| Bhattacharjee et al. | Recent advances in sensor-based detection of toxic dyes for bioremediation application: a review | |
| Wang et al. | Magnetic nanomotor-based maneuverable SERS probe | |
| CN103616367A (en) | Double-ion response type SERS (Surface Enhanced Raman Scattering) probe and preparation method thereof | |
| CN106290514A (en) | A kind of TiO based on silicon phthalocyanine functionalization2it is situated between and sees the aflatoxin Optical Electro-Chemistry detection method of crystal | |
| Wu et al. | Fluorescent molecularly imprinted nanoparticles for selective and rapid detection of ciprofloxacin in aquaculture water | |
| Gaikwad et al. | Fluorescent carbon-dots thin film for fungal detection and bio-labeling applications | |
| Gholami et al. | A new nano biosensor for maitotoxin with high sensitivity and selectivity based fluorescence resonance energy transfer between carbon quantum dots and gold nanoparticles | |
| Zhang et al. | Chemiluminescence chitosan hydrogels based on the luminol analog L-012 for highly sensitive detection of ROS | |
| CN105784666A (en) | Nano fluorescence biosensor as well as preparation method and application thereof | |
| Cao et al. | Zwitterionic electrochemiluminescence biointerface contributes to label‐free monitoring of exosomes dynamics in a fluidic microreaction device | |
| Rayappa et al. | Advances and effectiveness of metal–organic framework based bio/chemical sensors for rapid and ultrasensitive probing of antibiotic residues in foods |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210120 Address after: 405-47n, floor 4, building 3, No. 133, Yanling Road, Tianhe District, Guangzhou, Guangdong 510000 (office only) Patentee after: Guangzhou Chenze Intellectual Property Service Co.,Ltd. Address before: 266000 Qingdao University of Science & Technology, 99 Songling Road, Laoshan District, Qingdao, Shandong Patentee before: Qingdao University Of Science And Technology |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210308 Address after: 603, 6th floor, building D, mingrun business building, 388 Guangyun Road, Helong street, Baiyun District, Guangzhou, Guangdong 510440 (self declaration) Patentee after: Guangdong Youzhi Testing Technology Co.,Ltd. Address before: 405-47n, floor 4, building 3, No. 133, Yanling Road, Tianhe District, Guangzhou, Guangdong 510000 (office only) Patentee before: Guangzhou Chenze Intellectual Property Service Co.,Ltd. |
|
| TR01 | Transfer of patent right |