CN108459066A - Photoelectrochemistry biological immunosensor, preparation method thereof and application of photoelectrochemistry biological immunosensor in detection of specific base sequence - Google Patents
Photoelectrochemistry biological immunosensor, preparation method thereof and application of photoelectrochemistry biological immunosensor in detection of specific base sequence Download PDFInfo
- Publication number
- CN108459066A CN108459066A CN201810233839.8A CN201810233839A CN108459066A CN 108459066 A CN108459066 A CN 108459066A CN 201810233839 A CN201810233839 A CN 201810233839A CN 108459066 A CN108459066 A CN 108459066A
- Authority
- CN
- China
- Prior art keywords
- sensing device
- concentration
- added dropwise
- biological immune
- immune sensing
- 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
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 238000001514 detection method Methods 0.000 title abstract description 27
- 238000000034 method Methods 0.000 claims abstract description 11
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 52
- 230000005518 electrochemistry Effects 0.000 claims description 38
- 230000003287 optical effect Effects 0.000 claims description 38
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 32
- 239000000243 solution Substances 0.000 claims description 25
- 229910052982 molybdenum disulfide Inorganic materials 0.000 claims description 24
- 238000001035 drying Methods 0.000 claims description 23
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 23
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 22
- 108020004414 DNA Proteins 0.000 claims description 20
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 238000012360 testing method Methods 0.000 claims description 18
- 229910052980 cadmium sulfide Inorganic materials 0.000 claims description 17
- 229910052757 nitrogen Inorganic materials 0.000 claims description 16
- 238000006243 chemical reaction Methods 0.000 claims description 14
- CWQXQMHSOZUFJS-UHFFFAOYSA-N molybdenum disulfide Chemical compound S=[Mo]=S CWQXQMHSOZUFJS-UHFFFAOYSA-N 0.000 claims description 14
- 230000003321 amplification Effects 0.000 claims description 13
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 13
- 239000007788 liquid Substances 0.000 claims description 12
- 230000005622 photoelectricity Effects 0.000 claims description 12
- 239000002904 solvent Substances 0.000 claims description 12
- 239000003643 water by type Substances 0.000 claims description 12
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 11
- 239000000872 buffer Substances 0.000 claims description 11
- 239000006185 dispersion Substances 0.000 claims description 11
- 238000005406 washing Methods 0.000 claims description 11
- WUPHOULIZUERAE-UHFFFAOYSA-N 3-(oxolan-2-yl)propanoic acid Chemical compound OC(=O)CCC1CCCO1 WUPHOULIZUERAE-UHFFFAOYSA-N 0.000 claims description 10
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 10
- FTCIOUYXOOXMBV-UHFFFAOYSA-N OC(=O)c1ccccc1C(O)=O.C=CC(=O)OCCOCCOC(=O)C=C Chemical compound OC(=O)c1ccccc1C(O)=O.C=CC(=O)OCCOCCOC(=O)C=C FTCIOUYXOOXMBV-UHFFFAOYSA-N 0.000 claims description 10
- 229940116367 cadmium sulfide Drugs 0.000 claims description 10
- UHYPYGJEEGLRJD-UHFFFAOYSA-N cadmium(2+);selenium(2-) Chemical class [Se-2].[Cd+2] UHYPYGJEEGLRJD-UHFFFAOYSA-N 0.000 claims description 10
- 239000002096 quantum dot Substances 0.000 claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 239000002114 nanocomposite Substances 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 9
- 239000011734 sodium Substances 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- 239000006228 supernatant Substances 0.000 claims description 9
- 238000010792 warming Methods 0.000 claims description 9
- 239000011534 wash buffer Substances 0.000 claims description 9
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 claims description 8
- 239000011521 glass Substances 0.000 claims description 8
- 238000005119 centrifugation Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 239000008363 phosphate buffer Substances 0.000 claims description 6
- 229920001343 polytetrafluoroethylene Polymers 0.000 claims description 6
- 239000004810 polytetrafluoroethylene Substances 0.000 claims description 6
- 238000004062 sedimentation Methods 0.000 claims description 6
- 239000007787 solid Substances 0.000 claims description 6
- 229910052724 xenon Inorganic materials 0.000 claims description 6
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 claims description 6
- DKIDEFUBRARXTE-UHFFFAOYSA-N 3-mercaptopropanoic acid Chemical compound OC(=O)CCS DKIDEFUBRARXTE-UHFFFAOYSA-N 0.000 claims description 5
- 239000002253 acid Substances 0.000 claims description 5
- 229960005070 ascorbic acid Drugs 0.000 claims description 5
- 229910052793 cadmium Inorganic materials 0.000 claims description 5
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 claims description 5
- 238000011534 incubation Methods 0.000 claims description 5
- 239000000463 material Substances 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 claims description 4
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 claims description 4
- 239000002211 L-ascorbic acid Substances 0.000 claims description 4
- 235000000069 L-ascorbic acid Nutrition 0.000 claims description 4
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 4
- 239000000758 substrate Substances 0.000 claims description 4
- 238000002604 ultrasonography Methods 0.000 claims description 4
- 239000004201 L-cysteine Substances 0.000 claims description 3
- 235000013878 L-cysteine Nutrition 0.000 claims description 3
- 229920000877 Melamine resin Polymers 0.000 claims description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical class CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 3
- 229910019142 PO4 Inorganic materials 0.000 claims description 3
- 229910021607 Silver chloride Inorganic materials 0.000 claims description 3
- 235000018660 ammonium molybdate Nutrition 0.000 claims description 3
- ZSIAUFGUXNUGDI-UHFFFAOYSA-N hexan-1-ol Chemical class CCCCCCO ZSIAUFGUXNUGDI-UHFFFAOYSA-N 0.000 claims description 3
- 150000007974 melamines Chemical class 0.000 claims description 3
- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 239000004570 mortar (masonry) Substances 0.000 claims description 3
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 3
- 239000010452 phosphate Substances 0.000 claims description 3
- XNGIFLGASWRNHJ-UHFFFAOYSA-N phthalic acid Chemical compound OC(=O)C1=CC=CC=C1C(O)=O XNGIFLGASWRNHJ-UHFFFAOYSA-N 0.000 claims description 3
- -1 polytetrafluoroethylene Polymers 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 239000013049 sediment Substances 0.000 claims description 3
- 238000000527 sonication Methods 0.000 claims description 3
- 238000012546 transfer Methods 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- YKYOUMDCQGMQQO-UHFFFAOYSA-L Cadmium chloride Inorganic materials Cl[Cd]Cl YKYOUMDCQGMQQO-UHFFFAOYSA-L 0.000 claims description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 2
- QXDDUAGHAWNZHJ-UHFFFAOYSA-N [C].[Mo](=S)=S Chemical compound [C].[Mo](=S)=S QXDDUAGHAWNZHJ-UHFFFAOYSA-N 0.000 claims description 2
- 230000005611 electricity Effects 0.000 claims description 2
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 claims 3
- NIXOWILDQLNWCW-UHFFFAOYSA-M Acrylate Chemical compound [O-]C(=O)C=C NIXOWILDQLNWCW-UHFFFAOYSA-M 0.000 claims 1
- 150000001412 amines Chemical class 0.000 claims 1
- 238000007664 blowing Methods 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 238000007689 inspection Methods 0.000 claims 1
- 238000010926 purge Methods 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 239000002585 base Substances 0.000 description 33
- 208000025113 myeloid leukemia Diseases 0.000 description 7
- 229910052961 molybdenite Inorganic materials 0.000 description 6
- 210000003887 myelocyte Anatomy 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 239000001257 hydrogen Substances 0.000 description 3
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N titanium dioxide Inorganic materials O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 description 3
- 208000019838 Blood disease Diseases 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 2
- 239000005864 Sulphur Substances 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 229910021389 graphene Inorganic materials 0.000 description 2
- 208000014951 hematologic disease Diseases 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 238000012544 monitoring process Methods 0.000 description 2
- 239000002105 nanoparticle Substances 0.000 description 2
- 230000001699 photocatalysis Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- YUKQRDCYNOVPGJ-UHFFFAOYSA-N thioacetamide Chemical class CC(N)=S YUKQRDCYNOVPGJ-UHFFFAOYSA-N 0.000 description 2
- GDSOZVZXVXTJMI-SNAWJCMRSA-N (e)-1-methylbut-1-ene-1,2,4-tricarboxylic acid Chemical compound OC(=O)C(/C)=C(C(O)=O)\CCC(O)=O GDSOZVZXVXTJMI-SNAWJCMRSA-N 0.000 description 1
- ZOKXTWBITQBERF-UHFFFAOYSA-N Molybdenum Chemical compound [Mo] ZOKXTWBITQBERF-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003139 buffering effect Effects 0.000 description 1
- CJOBVZJTOIVNNF-UHFFFAOYSA-N cadmium sulfide Chemical compound [Cd]=S CJOBVZJTOIVNNF-UHFFFAOYSA-N 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 208000024207 chronic leukemia Diseases 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000003426 co-catalyst Substances 0.000 description 1
- 239000002131 composite material Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 208000018706 hematopoietic system disease Diseases 0.000 description 1
- 238000000703 high-speed centrifugation Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052750 molybdenum Inorganic materials 0.000 description 1
- 239000011733 molybdenum Substances 0.000 description 1
- 230000001613 neoplastic effect Effects 0.000 description 1
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 239000011941 photocatalyst Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- DLFVBJFMPXGRIB-UHFFFAOYSA-N thioacetamide Natural products CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
- G01N27/3278—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction involving nanosized elements, e.g. nanogaps or nanoparticles
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Physics & Mathematics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Electrochemistry (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Engineering & Computer Science (AREA)
- Nanotechnology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a photoelectrochemistry biological immunosensor, a preparation method thereof and application of the photoelectrochemistry biological immunosensor to detection of a specific base sequence. The invention takes the electrochemical signal as the detection basis, and simply and rapidly detects the concentration of the specific base sequence by detecting the magnitude of the bright and dark current on the surface of the electrode, thereby determining the concentration of the detected object. The method has the advantages of simple operation, strong specificity and high sensitivity.
Description
Technical field
The present invention relates to a kind of biological immune sensing device, specifically a kind of optical electro-chemistry biological immune sensing device and its
The application of preparation method and detection specific base sequence.
Background technology
G-C is found for the first time from Wang in 2009 etc.3N4Since water hydrogen manufacturing can be catalytically decomposed under visible light illumination, g-C3N4
Rapidly become research hotspot (Wang X, Maeda k, ThomasA, the etal.A metal-free of worldwide photoelectric field
polymeric photo-catalyst for hydrogen production from water under visible
light[J].Nat Mater,2009.8：76.).
Xiang and its project group membership will be layered the MoS of shape by solvent-thermal method2/ graphene composite catalyst supports
In TiO2On nano particle, Photocatalyzed Hydrogen Production efficiency significantly improves (XIANG Q, YU J, JARONIEC M.Synergetic
effect of MoS2and graphene as cocatalysts for enhanced photocatalytic
H2production activity of TiO2 nanoparticles[J].Journal of the American
Chemical Society,2012,134(15):6575-6578.)。
Zong et al. thinks MoS2With CdS due to identical S2-Ion, hetero-junctions between the two are more readily formed, pass through
On CdS but carry co-catalyst MoS2Successfully improve CdS photoelectric properties (ZONG X, YAN H, WU G,
etal.ENhancement of photocatalytic H2evolution on CdS by loading MoS2as
cocalyst under visible light irradiation[J].Journal of the American Chemical
Society,2008,130(23):7176-7177.)。
The neoplastic hematologic disorder that people's chronic myelocytic leukemia is a kind of to be generated because of specific chromosomal exception, in China
Account for about the 70% of chronic leukemia.Chromosome is made of a succession of gene, each gene is responsible for certain special work(
Can, the change of base sequence can cause normal gene composition to change, and then chromosome can be caused to be abnormal, to generate disease
Become.Therefore, the change of monitoring gene composition has great meaning to early detection and treatment people's chronic myelocytic leukemia.
Optical electro-chemistry be exactly under light illumination, when inorganic semiconductor material by energy be more than its energy gap when, electronics
Conduction band is transitted to from valence band, hole is generated in valence band, electronics, i.e. electron-hole pair are generated on conduction band, this is to electron hole pair
One kind may be again it is compound, it is alternatively possible to be in electronics transfer to the electron acceptor in external circuit or solution on conduction band,
Cause the generation of photoelectric current.
The method of present invention optical electro-chemistry can carry out the specific base of people's chronic myelocytic leukemia quantitative
Detection, to reach monitoring gene lesion, and then the purpose treated earlier.
Invention content
The present invention is intended to provide a kind of optical electro-chemistry biological immune sensing device and preparation method thereof and detection particular bases sequence
The application of row.The present invention is realized pair and the relevant specific alkali of people's myelocytic leukemia by building optical electro-chemistry immunosensor
The detection of base, method is simple, high specificity, high sensitivity.
Optical electro-chemistry biological immune sensing device of the present invention is to pass through layer assembly using FTO conductive glass electrodes as substrate
Method is in the surface of substrate covering phthalic acid diethylene glycol diacrylate, thin layer carbonitride and thin layer molybdenum disulfide
Mixture and cadmium sulfide quantum dot nano composite material (PDDA/C3N4+MoS2/ CdS QDs), in the nano combined material
The surface of material is fixed with basic base sequence by S-Cd keys.Optical electro-chemistry biological immune sensing device of the present invention is with electrochemical signals
For detection basis, by the detection to electrode surface light and shade size of current, simple and quick detects the dense of specific base sequence
Degree, and then determine the concentration of measured object.
The basis base sequence is 5 '-CAA-AAG-CCC-TTC-TTT-C3-SH, the basis base sequence meeting and target
DNA sequence dna occurs specific base complementation and combines.
The preparation method of optical electro-chemistry biological immune sensing device of the present invention, includes the following steps：
Step 1：It weighs 1.402g melamines to be placed in crucible, be then placed in Muffle furnace, be warming up to 550 DEG C and keep the temperature
4 hours (heating rate is 3 DEG C/min), crucible is taken out after heat preservation, is cooled to room temperature, gained sample is filled in mortar
Point grind into powder, then powder is transferred in Muffle furnace, be warming up to 500 DEG C and keep the temperature 2 hours (heating rate is 5 DEG C/
Min), gained sample is dispersed in water with the concentration of 1mg/mL after cooling, ultrasound 4 hours, then with the rotating speed of 9000r/min
10min is centrifuged, supernatant liquor is poured out, will be added in secondary water after the sediment vacuum drying after centrifugation, obtain a concentration of 1mg/
The thin layer carbonitride dispersion liquid of mL.
Step 2：At room temperature, 40mg ammonium molybdates and 80mg thiocarbamides are dissolved in 25ml dimethylformamides, are uniformly mixed
After be transferred in polytetrafluoroethylene (PTFE) autoclave, reacted 24 hours at 200 DEG C, you can obtain the molybdenum disulfide solution of black,
It is lyophilized at -60 DEG C after high speed centrifugation, gained molybdenum disulfide solid is dispersed in the concentration of 1mg/mL in secondary water, it is spare；
Step 3：Weigh 0.1467g CdCl2In three-necked flask, then 40mL secondary waters are added thereto, stirring and dissolving is complete
Entirely, the mercaptopropionic acid (MPA) of 172 μ L is then added dropwise, leads to nitrogen, adjusts pH to 11 with the NaOH of 1mol/L, is then added dropwise
The mixed solution of 0.0601g thioacetamides (TAA) and 10mL secondary waters, be warming up to 80 DEG C reaction 2h, cooled to room temperature,
Isometric absolute ethyl alcohol sedimentation is added into reaction solution, centrifuges (rotating speed 8000r/min, 15min), supernatant is poured out, with two
Volume constant volume is 8mL by secondary water, you can obtains water-soluble cadmiumsulfide quantum dot solution (CdS QDs)；
Step 4：The thin layer carbonitride dispersion liquid of a concentration of 1mg/mL that step 1 obtains and step 2 are obtained a concentration of
The molybdenum disulfide dispersion liquid of 1mg/mL by volume 1:The ratio of 1-3 mixes, and oscillator shakes to being uniformly mixed, and obtains thin layer nitrogen
Change carbon-molybdenum disulfide mixed liquor, use of purchasing is transferred in 4 DEG C；
Step 5：FTO conductive glass electrodes are subjected to ultrasound with secondary water, acetone, absolute ethyl alcohol successively with Sonication
Cleaning, each 3 times, 15 minutes every time, is subsequently placed in 60 DEG C of baking ovens 8~12 hours, spare；
Step 6：The FTO conductive glass electrodes cleaned through step 5 are tested out into conducting surface with multimeter, area is fixed as
5*9mm2, 30uL phthalic acid diethylene glycol diacrylate (PDDA) is added dropwise on conducting surface, room temperature stands 1 hour, uses
10mM, the phosphoric acid buffer solvent washing that pH value is 7.4, nitrogen drying；Then in phthalic acid diethylene glycol diacrylate
Surface thin layer carbonitride-molybdenum disulfide mixed liquor that 20-25uL steps 4 obtain is added dropwise, dry under natural conditions；Then it drips again
30uL phthalic acid diethylene glycol diacrylate (PDDA), room temperature is added to stand 1 hour, with 100mM, the phosphorus that pH value is 7.4
Acid buffering solvent washing, nitrogen drying；The water-soluble cadmiumsulfide quantum dot solution of 15-20uL steps 3 acquisition, naturally dry are added dropwise again
It is rinsed afterwards with secondary water, N2Drying, you can obtain nanocomposite modified electrode；
Step 7：By 1 μM of basic base sequence and 10mM tricarboxylics methyl acid phosphate (TCEP) by volume 100:1 ratio
Mixing, room temperature stands activation 1 hour after fully shaking, with disconnect disulfide bond (sulfydryl in basic base sequence easily with itself
Sulfydryl forms disulfide bond), the basis after 30uL is activated then is added dropwise in the surface of step 6 gained nanocomposite modified electrode
Base sequence is subsequently placed at 4 DEG C and is protected from light 12 hours of incubation；Electrode uses pH value after taking out be 8.0 TE wash buffers, N2
Drying, then six sulfydryls hexanols (MCH) of 10mM, 30uL are added dropwise and carry out the cadmium of enclosed-electrode surface exposure (with it in electrode surface
Formed sulphur cadmium key carry out enclosed-electrode), be subsequently placed in water-bath constant temperature oscillator and be incubated 1 hour at 37 DEG C, after taking-up use 10mM,
The PBS phosphoric acid buffer solvent washings that pH value is 7.4, N2Drying, obtains optical electro-chemistry biological immune sensing device.
The application of optical electro-chemistry biological immune sensing device of the present invention is white for detecting chronic myelocytic as detection reagent
The concentration of blood disease particular bases, specifically comprises the following steps：
Step 8：The target dna of 30uL concentration to be measured is added dropwise on the surface of optical electro-chemistry biological immune sensing device of the present invention,
It is incubated 90-100 minutes at 37 DEG C, the TE wash buffers for being 8.0 with pH value after taking-up, N2Drying；Then 30uL letters are added dropwise
Number amplification factor, and be incubated 110-120 minutes at 37 DEG C, the phosphoric acid buffer solvent washing for being 8.0 with pH after taking-up, nitrogen is blown
It does to get biological immune sensing device to be measured；
Step 9：By biological immune sensing device to be measured that step 8 obtains 2.5mL contain 0.1mol/L ascorbic acid and
Optical electro-chemistry test is carried out in 0.1mol/L, the phosphate buffer that pH value is 7.4, obtains photoelectricity figure；Using in gained photoelectricity figure
Difference between light and shade electric current obtains the concentration of object to be measured DNA with the standard relationship curve of object to be measured DNA concentration.
In step 8, the signal amplification factor is the CdSe quantum dots (CdSe QDs) of carboxylated, preparation method packet
Include following steps：
8a, 1.89g Na are weighed2SO3It is added in three-necked flask with 0.395g Se powder, 50ml secondary waters is then added, 70
It is stirred at reflux reaction 8 hours at~80 DEG C, obtains the Na of 0.1mol/L2SeSO3Solution.
8b, 0.2510g CdCl are weighed2In three-necked flask, 200ml secondary waters are added, are added after stirring and dissolving
The L-cysteine (L-cys) of 2.64mL, 1mol/L, it is 11 that 1mol/L sodium hydroxide solution tune pH value is used after stirring evenly, and is held
The Na of 0.1mol/L made from 0.75mL steps 1a is added after continuous logical nitrogen half an hour2SeSO3It is small to rise to 80 DEG C of reactions 2 for solution
When, isometric absolute ethyl alcohol sedimentation is added into reaction solution, centrifuges (rotating speed 8000r/min, 15min), pours out for natural cooling
Solid is settled to 15mL, you can obtain signal amplification factor by supernatant with secondary water --- the CdSe quantum dots of carboxylated
(CdSe QDs)。
In step 9, when carrying out optical electro-chemistry test, using three-electrode system, using Ag-AgCl electrodes as reference electrode, Pt
Silk electrode be auxiliary electrode, biological immune sensing device to be measured be working electrode, test using xenon lamp as light source, lamp cap with it is interelectrode
Distance is fixed as 2.5cm, and xenon lamp irradiation time is 20 seconds, and it is 20 seconds to be protected from light the time also, and the testing time is 400 seconds, is finally obtained
Photoelectricity figure.
In step 9, the standard relationship curve is to obtain by the following method：
30uL target dna standard samples are added dropwise on the surface of optical electro-chemistry biological immune sensing device of the present invention, at 37 DEG C
It is incubated 100 minutes, the TE wash buffers for being 8.0 with pH value after taking-up, N2Drying；Then 30uL signal amplification factors are added dropwise,
Up to standard sample biological immune sensing device；Gained standard sample biological immune sensing device is contained 0.1mol/L in 2.5mL to resist
Optical electro-chemistry test is carried out in bad hematic acid and 0.1mol/L, the phosphate buffer that pH value is 7.4, obtain photoelectricity figure (see Fig. 2)；With
The logarithm of target dna (myelocytic leukemia particular bases) concentration is abscissa, is fitted using light and shade current difference as ordinate,
It can be obtained standard relationship curve (Fig. 2 illustrations).Detection shows the concentration when the particular bases of myelocytic leukemia in 2uM-1pM
Concentration range in detection line in a linear relationship reach 1pM.
The concentration of the target dna standard sample is respectively 2uM, 100nM, 10nM, 1nM, 0.1nM, 0.01nM, 1pM.
The present invention uses the MoS of two-dimensional structure2With the g-C of class graphene-structured3Bases of the N4 as biological immune sensing device
Bottom.By MoS2With C3N4In conjunction with easily forming heterojunction structure so that photoinduction g-C3N4The electronics that surface generates can move to rapidly
MoS2In nanometer sheet, the distance of charge transmission is shortened, photoelectric efficiency is improved.Under the irradiation of light, electrode surface generates electricity
Stream, we collect current signal, and the concentration and biological immune of base sequence are judged according to the size of photo-signal
The specificity of sensor judges the accuracy of base sequence.
Compared with the prior art, beneficial effects of the present invention are embodied in：
1, the present invention by build optical electro-chemistry immunosensor realize it is pair relevant specific with people's myelocytic leukemia
The detection of base, method is simple, high specificity, high sensitivity.
2, sample size needed for the detection in the present invention pair with the relevant particular bases of people's myelocytic leukemia is few, testing cost
It is low.
3, the present invention is by building phthalic acid diethylene glycol diacrylate/thin layer carbonitride and thin layer molybdenum disulfide
Mixture/cadmiumsulfide quantum dot complexes membrane prepare biological immune sensing device, photocurrent variations are apparent after concentration changes, raw
Object compatibility is good, and has very high stability.
Description of the drawings
Fig. 1 is preparation and the detection process principle schematic of optical electro-chemistry biological immune sensing device of the present invention.
Fig. 2 present invention is respectively 2uM, 100nM, 10nM, 1nM, 0.1nM, 0.01nM, 1pM (from left to right) to concentration
Target DNA concentration carries out the test result of Photoelectric Detection, and illustration is standard curve relationship.
Fig. 3 is scanning electron microscope (SEM) table of the mixture of thin layer carbonitride and thin layer molybdenum disulfide in the present invention
The result of sign.
Fig. 4 is the result of transmission electron microscope (TEM) characterization of cadmiumsulfide quantum dot compound in the present invention.
Specific implementation mode
Embodiment 1：
The preparation method of optical electro-chemistry biological immune sensing device is as follows in the present embodiment：
1,1.402g melamines are weighed to be placed in crucible, are then placed in Muffle furnace, 550 DEG C is warming up to and heat preservation 4 is small
When (heating rate be 3 DEG C/min), crucible is taken out after heat preservation, is cooled to room temperature, in mortar fully by gained sample
Grind into powder, then powder is transferred in Muffle furnace, be warming up to 500 DEG C and keeps the temperature 2 hours (heating rate is 5 DEG C/min),
Gained sample is dispersed in water with the concentration of 1mg/mL after cooling, then ultrasound 4 hours is centrifuged with the rotating speed of 9000r/min
10min pours out supernatant liquor, will be added in secondary water after the sediment vacuum drying after centrifugation, obtains a concentration of 1mg/mL's
Thin layer carbonitride dispersion liquid.
2, at room temperature, 40mg ammonium molybdates and 80mg thiocarbamides are dissolved in 25ml dimethylformamides, are turned after mixing
It moves in polytetrafluoroethylene (PTFE) autoclave, is reacted 24 hours at 200 DEG C, you can obtain the molybdenum disulfide solution of black, high speed
It is lyophilized at -60 DEG C after centrifugation, gained molybdenum disulfide solid is dispersed in the concentration of 1mg/mL in secondary water, it is spare；
3,0.1467g CdCl are weighed2In three-necked flask, then 40mL secondary waters being added thereto, stirring and dissolving is complete,
Then the mercaptopropionic acid (MPA) of 172 μ L is added dropwise, leads to nitrogen, adjusts pH to 11 with the NaOH of 1mol/L, 0.0601g is then added dropwise
The mixed solution of thioacetamide (TAA) and 10mL secondary waters are warming up to 80 DEG C of reaction 2h, cooled to room temperature, to reaction
Isometric absolute ethyl alcohol sedimentation is added in liquid, centrifuges (rotating speed 8000r/min, 15min), pours out supernatant, it will with secondary water
Volume constant volume is 8mL, you can obtains water-soluble cadmiumsulfide quantum dot solution (CdS QDs)；
4, a concentration of 1mg/ for obtaining the thin layer carbonitride dispersion liquid of a concentration of 1mg/mL that step 1 obtains and step 2
The molybdenum disulfide dispersion liquid of mL by volume 1:3 ratio mixing, oscillator shake to being uniformly mixed, and obtain thin layer carbonitride-
Molybdenum disulfide mixed liquor transfers use of purchasing in 4 DEG C；
5, FTO conductive glass electrodes are carried out with secondary water, acetone, absolute ethyl alcohol successively with Sonication ultrasonic clear
It washes, each 3 times, 15 minutes every time, is subsequently placed in 60 DEG C of baking ovens 8~12 hours, it is spare；
6, the FTO conductive glass electrodes cleaned through step 5 are tested out into conducting surface with multimeter, area is fixed as 5*
9mm2, 30uL phthalic acid diethylene glycol diacrylate (PDDA) is added dropwise on conducting surface, room temperature stands 1 hour, uses
10mM, the phosphoric acid buffer solvent washing that pH value is 7.4, nitrogen drying；Then in phthalic acid diethylene glycol diacrylate
Surface thin layer carbonitride-molybdenum disulfide mixed liquor that 20uL steps 4 obtain is added dropwise, dry under natural conditions；Then it is added dropwise again
30uL phthalic acid diethylene glycol diacrylate (PDDA), room temperature stand 1 hour, with 100mM, the phosphoric acid that pH value is 7.4
Buffer solvent rinses, nitrogen drying；The water-soluble cadmiumsulfide quantum dot solution of 15uL steps 3 acquisition is added dropwise again, uses after natural drying
Secondary water is rinsed, N2Drying, you can obtain nanocomposite modified electrode；
7, by 1 μM of basic base sequence and 10mM tricarboxylics methyl acid phosphate (TCEP) by volume 100:1 ratio mixing,
Room temperature stands activation 1 hour after fully shaking, with disconnect disulfide bond (sulfydryl in basic base sequence easily with the sulfydryl shape of itself
At disulfide bond), the basic base sequence after 30uL is activated then is added dropwise in the surface of step 8 gained nanocomposite modified electrode
Row are subsequently placed at 4 DEG C and are protected from light 12 hours of incubation；Electrode uses pH value after taking out be 8.0 TE wash buffers, N2Drying,
Six sulfydryls hexanols (MCH) of 10mM, 30uL are added dropwise to cadmium (the formed sulphur for carrying out enclosed-electrode surface exposure in electrode surface again
Cadmium key carrys out enclosed-electrode), it is subsequently placed in water-bath constant temperature oscillator and is incubated 1 hour at 37 DEG C, be with 10mM, pH value after taking-up
7.4 PBS phosphoric acid buffer solvent washings, N2Drying, obtains optical electro-chemistry biological immune sensing device.
Using optical electro-chemistry biological immune sensing device manufactured in the present embodiment as detection reagent for detecting chronic myelocytic
The concentration of leukaemia particular bases, it is specific as follows：
8, the target dna of 30uL concentration to be measured is added dropwise on the surface of optical electro-chemistry biological immune sensing device, is incubated at 37 DEG C
It educates 90-100 minutes, the TE wash buffers for being 8.0 with pH value after taking-up, N2Drying；Then 30uL signal amplification factors are added dropwise,
And be incubated 120 minutes at 37 DEG C, the phosphoric acid buffer solvent washing for being 8.0 with pH after taking-up, nitrogen dries up to get biology to be measured
Immunosensor；
9, the biological immune sensing device to be measured that step 8 obtains is contained into 0.1mol/L ascorbic acid and 0.1mol/ in 2.5mL
L, optical electro-chemistry test is carried out in the phosphate buffer that pH value is 7.4, obtains photoelectricity figure；Utilize light and shade electric current in gained photoelectricity figure
Between the standard relationship curve of difference and object to be measured DNA concentration obtain the concentration of object to be measured DNA.
In step 8, the signal amplification factor is the CdSe quantum dots (CdSe QDs) of carboxylated, preparation method packet
Include following steps：
8a, 1.89g Na are weighed2SO3It is added in three-necked flask with 0.395g Se powder, 50ml secondary waters is then added, 70
It is stirred at reflux reaction 8 hours at~80 DEG C, obtains the Na of 0.1mol/L2SeSO3Solution.
8b, 0.2510g CdCl are weighed2In three-necked flask, 200ml secondary waters are added, are added after stirring and dissolving
The L-cysteine (L-cys) of 2.64mL, 1mol/L, it is 11 that 1mol/L sodium hydroxide solution tune pH value is used after stirring evenly, and is held
The Na of 0.1mol/L made from 0.75mL steps 1a is added after continuous logical nitrogen half an hour2SeSO3It is small to rise to 80 DEG C of reactions 2 for solution
When, isometric absolute ethyl alcohol sedimentation is added into reaction solution, centrifuges (rotating speed 8000r/min, 15min), pours out for natural cooling
Solid is settled to 15mL, you can obtain signal amplification factor by supernatant with secondary water --- the CdSe quantum dots of carboxylated
(CdSe QDs)。
In step 9, when carrying out optical electro-chemistry test, using three-electrode system, using Ag-AgCl electrodes as reference electrode, Pt
Silk electrode be auxiliary electrode, biological immune sensing device to be measured be working electrode, test using xenon lamp as light source, lamp cap with it is interelectrode
Distance is fixed as 2.5cm, and xenon lamp irradiation time is 20 seconds, and it is 20 seconds to be protected from light the time also, and the testing time is 400 seconds, is finally obtained
Photoelectricity figure.
In step 9, the standard relationship curve is to obtain by the following method：
30uL target dna standard samples are added dropwise on the surface of optical electro-chemistry biological immune sensing device of the present invention, at 37 DEG C
It is incubated 90-100 minutes, the TE wash buffers for being 8.0 with pH value after taking-up, N2Drying；Then be added dropwise 30uL signals amplification because
Son is to get standard sample biological immune sensing device；Gained standard sample biological immune sensing device is contained into 0.1mol/L in 2.5mL
Optical electro-chemistry test is carried out in ascorbic acid and 0.1mol/L, the phosphate buffer that pH value is 7.4, obtain photoelectricity figure (see Fig. 2)；
Using the logarithm of target dna (myelocytic leukemia particular bases) concentration as abscissa, intended using light and shade current difference as ordinate
It closes, you can obtain standard relationship curve (Fig. 2 illustrations).Detection shows the concentration when the particular bases of myelocytic leukemia in 2uM-
Detection line in a linear relationship reaches 1pM in the concentration range of 1pM.
The concentration of the target dna standard sample is respectively 2uM, 100nM, 10nM, 1nM, 0.1nM, 0.01nM, 1pM.
The stability of optical electro-chemistry biological immune sensing device of the present invention is good, high specificity, high sensitivity, to NH2- DNA's
Concentration Testing range is 1PM-2 μM, and detection line reaches 1PM, has very high practicability.
Embodiment 2：
The preparation process of the present embodiment with embodiment 1, unlike in step 4, thin layer carbonitride dispersion liquid and curing
Molybdenum dispersion liquid by volume 1:1 ratio mixing.Gained biological immune sensing device and the pattern of embodiment 1 are similar with property, lead to
The detection to people's myelocyte specific base is crossed, it is found that result is identical.
Embodiment 3：
The preparation process of the present embodiment with embodiment 1, unlike in step 6, in O-phthalic acid diethylene glycol dipropyl
Thin layer carbonitride-molybdenum disulfide mixed liquor that 25uL steps 4 obtain is added dropwise in the surface of olefin(e) acid ester.Gained biological immune sensing device and
The pattern of embodiment 1 is similar with property, by the detection to people's myelocyte specific base, it is found that result is identical.
Embodiment 4：
With embodiment 1, the difference is that in step 6, the water-soluble sulphur that 20uL steps 3 obtain is added dropwise in the preparation process of the present embodiment
Cadmium quantum dot solution.Gained biological immune sensing device and the pattern of embodiment 1 are similar with property, by special to people's myelocyte
The detection of base finds that result is identical.
Embodiment 5：
The preparation process of the present embodiment is with embodiment 1, the difference is that in step 8, incubation time 100min is changed to 90min.
Gained biological immune sensing device and the pattern of embodiment 1 are similar with property, by the detection to people's myelocyte specific base, find
As a result identical.
Embodiment 6：
The preparation process of the present embodiment with embodiment 1, unlike in step 8, be added dropwise 30uL signal amplification factors, and
It is incubated 110 minutes at 37 DEG C.Gained biological immune sensing device and the pattern of embodiment 1 are similar with property, by people's myelocyte
The detection of specific base finds that result is identical.
Claims (9)
1. a kind of optical electro-chemistry biological immune sensing device, it is characterised in that：The optical electro-chemistry biological immune sensing device is with FTO
Conductive glass electrode is substrate, and O-phthalic acid diethylene glycol two is covered on the surface of the substrate by the method for layer assembly
Acrylate, the mixture of thin layer carbonitride and thin layer molybdenum disulfide and cadmium sulfide quantum dot nano composite material, described
The surface of nanocomposite is fixed with basic base sequence by S-Cd keys.
2. optical electro-chemistry biological immune sensing device according to claim 1, it is characterised in that：
The basis base sequence is 5 '-CAA-AAG-CCC-TTC-TTT-C3-SH, the basis base sequence meeting and target dna
Sequence occurs specific base complementation and combines.
3. a kind of preparation method of optical electro-chemistry biological immune sensing device as claimed in claim 1 or 2, it is characterised in that including such as
Lower step：
Step 1：It weighs 1.402g melamines to be placed in crucible, be then placed in Muffle furnace, be warming up to 550 DEG C and heat preservation 4 is small
When, crucible is taken out after heat preservation, is cooled to room temperature, by gained sample in mortar grind into powder, then powder is shifted
Into Muffle furnace, it is warming up to 500 DEG C and keeps the temperature 2 hours, be dispersed in water gained sample with the concentration of 1mg/mL after cooling, surpass
Sound 4 hours, pours out supernatant liquor after centrifugation, will be added in secondary water, obtain a concentration of after the sediment vacuum drying after centrifugation
The thin layer carbonitride dispersion liquid of 1mg/mL；
Step 2：At room temperature, 40mg ammonium molybdates and 80mg thiocarbamides are dissolved in 25ml dimethylformamides, are turned after mixing
It moves in polytetrafluoroethylene (PTFE) autoclave, is reacted 24 hours at 200 DEG C, you can obtain the molybdenum disulfide solution of black, centrifuge
It is lyophilized at -60 DEG C afterwards, gained molybdenum disulfide solid is dispersed in the concentration of 1mg/mL in secondary water, it is spare；
Step 3：Weigh 0.1467g CdCl2In three-necked flask, then 40mL secondary waters are added thereto, stirring and dissolving is complete, so
The mercaptopropionic acid of 172 μ L is added dropwise afterwards, leads to nitrogen, adjusts pH to 11 with the NaOH of 1mol/L, 0.0601g thioacetyls is then added dropwise
The mixed solution of amine and 10mL secondary waters, is warming up to 80 DEG C of reaction 2h, and cooled to room temperature is added isometric into reaction solution
Absolute ethyl alcohol sedimentation, supernatant is poured out in centrifugation, by volume constant volume is 8mL with secondary water, you can obtain water-soluble cadmium sulfide quantum
Point solution；
Step 4：A concentration of 1mg/ that the thin layer carbonitride dispersion liquid of a concentration of 1mg/mL that step 1 obtains and step 2 are obtained
The molybdenum disulfide dispersion liquid of mL by volume 1:The ratio of 1-3 mixes, and oscillator shakes to being uniformly mixed, and obtains thin layer nitridation
Carbon-molybdenum disulfide mixed liquor transfers use of purchasing in 4 DEG C；
Step 5：FTO conductive glass electrodes are subjected to ultrasound clearly with secondary water, acetone, absolute ethyl alcohol successively with Sonication
It washes, is subsequently placed in 60 DEG C of baking ovens 8~12 hours, it is spare；
Step 6：The FTO conductive glass electrodes cleaned through step 5 are tested out into conducting surface with multimeter, area is fixed as 5*
9mm2, 30uL phthalic acid diethylene glycol diacrylate is added dropwise on conducting surface, room temperature stands 1 hour, with 10mM, pH value
For 7.4 phosphoric acid buffer solvent washing, nitrogen drying；Then it is added dropwise on the surface of phthalic acid diethylene glycol diacrylate
Thin layer carbonitride-molybdenum disulfide mixed liquor that 20-25uL steps 4 obtain dries under natural conditions；Then 30uL neighbour's benzene is added dropwise again
Dicarboxylic omega-diol diacrylate, room temperature stand 1 hour, with 100mM, the phosphoric acid buffer solvent washing that pH value is 7.4, nitrogen
Air-blowing is dry；The water-soluble cadmiumsulfide quantum dot solution of 15-20uL steps 3 acquisition is added dropwise again, is rinsed after natural drying with secondary water, N2
Drying, you can obtain nanocomposite modified electrode；
Step 7：By 1 μM of basic base sequence and 10mM tricarboxylics methyl acid phosphate by volume 100:1 ratio mixing, fully shakes
It swings rear room temperature and stands activation 1 hour, to disconnect disulfide bond, then in the surface of step 6 gained nanocomposite modified electrode
The basic base sequence after 30uL activation is added dropwise, is subsequently placed at 4 DEG C and is protected from light 12 hours of incubation；It is with pH value after electrode taking-up
8.0 TE wash buffers, N2Drying, then six sulfydryls hexanols of 10mM, 30uL are added dropwise and carry out enclosed-electrode table in electrode surface
The exposed cadmium in face is subsequently placed in water-bath constant temperature oscillator and is incubated 1 hour at 37 DEG C, and it is 7.4 that 10mM, pH value are used after taking-up
PBS phosphoric acid buffer solvent washings, N2Drying, obtains optical electro-chemistry biological immune sensing device.
4. a kind of application of optical electro-chemistry biological immune sensing device as claimed in claim 1 or 2, it is characterised in that：It is as inspection
Test agent is used to detect the concentration of chronic myelocytic leukemia particular bases.
5. application according to claim 4, it is characterised in that include the following steps：
Step 8：The target dna of 30uL concentration to be measured is added dropwise on the surface of the optical electro-chemistry biological immune sensing device, in 37 DEG C
Lower incubation 90-100 minutes, the TE wash buffers for being 8.0 with pH value after taking-up, N2Drying；Then the amplification of 30uL signals is added dropwise
The factor, and be incubated 110-120 minutes at 37 DEG C, the phosphoric acid buffer solvent washing for being 8.0 with pH after taking-up, nitrogen drying, i.e.,
Obtain biological immune sensing device to be measured；
Step 9：The biological immune sensing device to be measured that step 8 obtains is contained into 0.1mol/L ascorbic acid and 0.1mol/ in 2.5mL
L, optical electro-chemistry test is carried out in the phosphate buffer that pH value is 7.4, obtains photoelectricity figure；Utilize light and shade electric current in gained photoelectricity figure
Between the standard relationship curve of difference and object to be measured DNA concentration obtain the concentration of object to be measured DNA.
6. application according to claim 5, it is characterised in that：
In step 8, the signal amplification factor is the CdSe quantum dots of carboxylated, and preparation method includes the following steps：
8a, 1.89g Na are weighed2SO3It is added in three-necked flask with 0.395g Se powder, 50ml secondary waters is then added, 70~80
It is stirred at reflux reaction 8 hours at DEG C, obtains the Na of 0.1mol/L2SeSO3Solution；
8b, 0.2510g CdCl are weighed2In three-necked flask, 200ml secondary waters are added, be added after stirring and dissolving 2.64mL,
The L-cysteine of 1mol/L, it is 11 that 1mol/L sodium hydroxide solution tune pH value is used after stirring evenly, and maintaining nitrogen purge is half small
When after the Na of 0.1mol/L made from 0.75mL steps 1a is added2SeSO3Solution rises to 80 DEG C and reacts 2 hours, natural cooling, to
Isometric absolute ethyl alcohol sedimentation is added in reaction solution, centrifugation pours out supernatant, solid is settled to 15mL with secondary water, i.e.,
Signal amplification factor can be obtained --- the CdSe quantum dots of carboxylated.
7. application according to claim 5, it is characterised in that：
In step 9, when carrying out optical electro-chemistry test, using three-electrode system, using Ag-AgCl electrodes as reference electrode, Pt electricity
Extremely auxiliary electrode, biological immune sensing device to be measured are working electrode, are tested using xenon lamp as light source, lamp cap and interelectrode distance
It is fixed as 2.5cm, xenon lamp irradiation time is 20 seconds, and it is 20 seconds to be protected from light the time also, and the testing time is 400 seconds, finally obtains photoelectricity
Figure.
8. application according to claim 5, it is characterised in that：
In step 9, the standard relationship curve is to obtain by the following method：
30uL target dna standard samples are added dropwise on the surface of the optical electro-chemistry biological immune sensing device, 100 are incubated at 37 DEG C
Minute, the TE wash buffers for being 8.0 with pH value after taking-up, N2Drying；Then 30uL signal amplification factors are added dropwise to get standard
Sample biological immune sensing device；By gained standard sample biological immune sensing device 2.5mL contain 0.1mol/L ascorbic acid and
Optical electro-chemistry test is carried out in 0.1mol/L, the phosphate buffer that pH value is 7.4, obtains photoelectricity figure；With pair of target DNA concentration
Number is abscissa, is fitted using light and shade current difference as ordinate, you can obtains standard relationship curve.
9. application according to claim 8, it is characterised in that：
A concentration of 2uM-1pM of the target dna standard sample at least takes six different point values.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810233839.8A CN108459066B (en) | 2018-03-21 | 2018-03-21 | Photoelectrochemistry biological immunosensor, preparation method thereof and application of photoelectrochemistry biological immunosensor in detection of specific base sequence |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810233839.8A CN108459066B (en) | 2018-03-21 | 2018-03-21 | Photoelectrochemistry biological immunosensor, preparation method thereof and application of photoelectrochemistry biological immunosensor in detection of specific base sequence |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108459066A true CN108459066A (en) | 2018-08-28 |
| CN108459066B CN108459066B (en) | 2020-06-05 |
Family
ID=63237398
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810233839.8A Active CN108459066B (en) | 2018-03-21 | 2018-03-21 | Photoelectrochemistry biological immunosensor, preparation method thereof and application of photoelectrochemistry biological immunosensor in detection of specific base sequence |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108459066B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110927238A (en) * | 2019-12-12 | 2020-03-27 | 山东理工大学 | Preparation method and application of sandwich type photoelectrochemical sensor for detecting prostate specific antigen |
| CN111272848A (en) * | 2020-03-06 | 2020-06-12 | 安徽大学 | High-sensitivity photoelectrochemical biosensor for detecting miRNA159c and preparation and detection methods thereof |
| CN114689670A (en) * | 2022-03-21 | 2022-07-01 | 合肥海关技术中心 | MOF-based electrochemical sensor and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1995386A (en) * | 2006-08-22 | 2007-07-11 | 上海复星医药（集团）股份有限公司 | BCR-ABL gene fluorescence quantitative RT-PCR primer and probe and reagent kit |
| CN102175729A (en) * | 2011-01-13 | 2011-09-07 | 青岛科技大学 | Photoinduced electrochemical sensor manufactured by assembling functional grapheme and nano particle layer by layer |
| CN104891997A (en) * | 2015-05-27 | 2015-09-09 | 青岛大学 | Preparation method of graphite phase carbon nitride/molybdenum sulfide composite material |
-
2018
- 2018-03-21 CN CN201810233839.8A patent/CN108459066B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1995386A (en) * | 2006-08-22 | 2007-07-11 | 上海复星医药（集团）股份有限公司 | BCR-ABL gene fluorescence quantitative RT-PCR primer and probe and reagent kit |
| CN102175729A (en) * | 2011-01-13 | 2011-09-07 | 青岛科技大学 | Photoinduced electrochemical sensor manufactured by assembling functional grapheme and nano particle layer by layer |
| CN104891997A (en) * | 2015-05-27 | 2015-09-09 | 青岛大学 | Preparation method of graphite phase carbon nitride/molybdenum sulfide composite material |
Non-Patent Citations (3)
| Title |
|---|
| JIAN SHU 等: "Current Advances in Quantum-Dots-Based Photoelectrochemical Immunoassays", 《CHEMISTRY-AN ASIAN JOURNAL》 * |
| QIAN LI 等: "High Efficiency Photocatalysis for Pollutant Degradation with MoS2/C3N4 Heterostructures", 《LANGMUIR》 * |
| 刘康丽 等: "基于原位形成p—n结的新型光电化学传感器检测Hg2+", 《分析测试学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110927238A (en) * | 2019-12-12 | 2020-03-27 | 山东理工大学 | Preparation method and application of sandwich type photoelectrochemical sensor for detecting prostate specific antigen |
| CN110927238B (en) * | 2019-12-12 | 2022-10-11 | 山东理工大学 | Preparation method and application of sandwich type photoelectrochemical sensor for detecting prostate specific antigen |
| CN111272848A (en) * | 2020-03-06 | 2020-06-12 | 安徽大学 | High-sensitivity photoelectrochemical biosensor for detecting miRNA159c and preparation and detection methods thereof |
| CN111272848B (en) * | 2020-03-06 | 2022-04-26 | 安徽大学 | High-sensitivity photoelectrochemical biosensor for detecting miRNA159c and preparation and detection methods thereof |
| CN114689670A (en) * | 2022-03-21 | 2022-07-01 | 合肥海关技术中心 | MOF-based electrochemical sensor and preparation method and application thereof |
| CN114689670B (en) * | 2022-03-21 | 2024-02-02 | 合肥海关技术中心 | MOF-based electrochemical sensor and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108459066B (en) | 2020-06-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Using reduced graphene oxide-Ca: CdSe nanocomposite to enhance photoelectrochemical activity of gold nanoparticles functionalized tungsten oxide for highly sensitive prostate specific antigen detection | |
| Mehta et al. | Graphene quantum dot modified screen printed immunosensor for the determination of parathion | |
| Zhou et al. | Ternary electrochemiluminescence nanostructure of Au nanoclusters as a highly efficient signal label for ultrasensitive detection of cancer biomarkers | |
| Lin et al. | Sensitive amperometric immunosensor for α-fetoprotein based on carbon nanotube/gold nanoparticle doped chitosan film | |
| Zhang et al. | Photoelectrochemical response of carbon dots (CDs) derived from chitosan and their use in electrochemical imaging | |
| Chen et al. | Efficient Z-Scheme heterostructure based on TiO2/Ti3C2Tx/Cu2O to boost photoelectrochemical response for ultrasensitive biosensing | |
| Yang et al. | Ultrasensitive “signal-on” electrochemiluminescence immunosensor for prostate-specific antigen detection based on novel nanoprobe and poly (indole-6-carboxylic acid)/flower-like Au nanocomposite | |
| Li et al. | Quenched electrochemiluminescence of Ag nanoparticles functionalized g-C3N4 by ferrocene for highly sensitive immunosensing | |
| Zuo et al. | An electrochemiluminescent sensor for dopamine detection based on a dual-molecule recognition strategy and polyaniline quenching | |
| Shi et al. | Aptasensor based on tripetalous cadmium sulfide-graphene electrochemiluminescence for the detection of carcinoembryonic antigen | |
| Liu et al. | Application of ZnO quantum dots dotted carbon nanotube for sensitive electrochemiluminescence immunoassay based on simply electrochemical reduced Pt/Au alloy and a disposable device | |
| Tian et al. | Amplified quenching of electrochemiluminescence from CdS sensitized TiO 2 nanotubes by CdTe–carbon nanotube composite for detection of prostate protein antigen in serum | |
| Wang et al. | Ultrasensitive photoelectrochemical immunosensor for insulin detection based on dual inhibition effect of CuS-SiO2 composite on CdS sensitized C-TiO2 | |
| CN108459066A (en) | Photoelectrochemistry biological immunosensor, preparation method thereof and application of photoelectrochemistry biological immunosensor in detection of specific base sequence | |
| CN110082415B (en) | Photoelectric chemical detection probe based on conjugated polymer nanoparticles and application thereof | |
| Liu et al. | A novel photoelectrochemical immunosensor by integration of nanobody and ZnO nanorods for sensitive detection of nucleoside diphosphatase kinase-A | |
| Sedghi et al. | Fabrication of non-enzymatic glucose sensor based on nanocomposite of MWCNTs-COOH-Poly (2-aminothiophenol)-Au NPs | |
| CN112683971B (en) | Chloramphenicol photoelectrochemical aptamer sensor and preparation method and application thereof | |
| Jiang et al. | Two-dimensional Ti2C MXene-induced photocurrent polarity switching photoelectrochemical biosensing platform for ultrasensitive and selective detection of soluble CD146 | |
| CN111830104B (en) | Photoelectrochemistry biosensor and preparation method and application thereof | |
| Xu et al. | Renewable photoelectrochemical cytosensing platform for rapid capture and detection of circulating tumor cells | |
| Zhao et al. | Monolayer rubrene functionalized graphene-based eletrochemiluminescence biosensor for serum cystatin C detection with immunorecognition-induced 3D DNA machine | |
| Kang et al. | A novel signal amplification strategy electrochemical immunosensor for ultra-sensitive determination of p53 protein | |
| Mazhabi et al. | A label-free aptamer-based cytosensor for specific cervical cancer HeLa cell recognition through a gC 3 N 4–AgI/ITO photoelectrode | |
| Wang et al. | A sensitive quenched electrochemiluminescent DNA sensor based on the catalytic activity of gold nanoparticle functionalized MoS 2 |
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 |