CN107064509B - Detect the preparation and application of the optical electro-chemistry immunosensor of carcinomebryonic antigen - Google Patents
Detect the preparation and application of the optical electro-chemistry immunosensor of carcinomebryonic antigen Download PDFInfo
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Abstract
The invention discloses a kind of preparations and application study of the optical electro-chemistry immunosensor of detection carcinomebryonic antigen.The zinc oxide nano rod nanometer sheet of growing three-dimensional layered structure on the tin dioxide transparent conductive glass of doping fluorine, the cadmiumsulfide quantum dot of a large amount of additive Mn is loaded using it and identifies that the aptamers of carcinomebryonic antigen and the label that hybridizes with aptamers have the/DNA probe of tellurium/cadmium selenide core shell quantum dot, the polynary sensitization structure based on zinc oxide is formed, realizes initial signal amplification；Make the space steric effect for the conjugate itself that the hybridization of DNA probe solution is detached from the sensibilization for the decrease that electrode surface generates and carcinomebryonic antigen is formed with aptamers in conjunction with carcinomebryonic antigen specific recognition aptamers, it realizes further signal amplification, and then realizes the Sensitive Detection to carcinomebryonic antigen.
Description
Technical field
The present invention relates to nano material technology, optical electro-chemistry signal detection technique and immunoassay detection technique fields, more
The specifically preparation and application of a kind of optical electro-chemistry immunosensor for Sensitive Detection carcinomebryonic antigen.
Background technology
As a kind of very important tumor markers, carcinomebryonic antigen is played the part of emphatically in the clinical diagnosis and treatment of cancer
The role wanted.In recent years, various analysis methods are developed for the detection of carcinomebryonic antigen, as fluorescence, electrochemistry, electrification
Learn the analysis methods such as luminous, colorimetric.Although the detection of carcinomebryonic antigen may be implemented in these methods, there are complicated for operation, clever for they
The shortcomings of sensitivity is low, time-consuming, of high cost.Photoelectrochemical assay method is as a kind of emerging analysis method, due to its sensitivity
The advantages that high, easy to operate, at low cost, causes extensive concern！Photoelectric activity material is the weight in photoelectrochemical assay method
Component part is wanted, the common structure that is sensitized is typically half that different narrow band gaps is sensitized based on the semi-conducting material of broad-band gap
Conductor material.The recombination rate for reducing sensitization structure electrical-hole pair, it is to improve photoelectricity to the absorption region of visible light to enhance it
The key of chemical analysis performance.
Common recognition element is antibody in traditional immunosensor, is typically to be generated by cell strain or animal,
Acquisition process is complex and high cost, and in addition its stability is poor, is easily influenced by high temperature, acid-base condition, these intrinsic defects
It is significantly limited to be widely applied.Therefore it is badly in need of seeking novel effective molecular recognition element.In addition, carcinomebryonic antigen is in people
The content of class serum is relatively low, develops effective signal amplification mode, the sensitivity of its analysis detection is improved, for clinical cancer
Analyzing and diagnosing has great importance.
Invention content
The purpose of the present invention is the oxidations of the growing three-dimensional layered structure on the tin dioxide transparent conductive glass of doping fluorine
Zinc nanometer rods-nanometer sheet, in the cadmiumsulfide quantum dot of a large amount of additive Mn of its area load, wherein the stannic oxide of doping fluorine is saturating
Bright electro-conductive glass is abbreviated as FTO, and zinc oxide nano rod-nanometer sheet of layered structure writes a Chinese character in simplified form ZnO NRs-NSs, the vulcanization of additive Mn
Cadmium quantum dot is abbreviated as CdS:Mn, using aptamers as the recognition element of carcinomebryonic antigen, 5 ' ends are marked with cadmium telluride/cadmium selenide
The DNA probe of core-shell quanta dots is captured to electrode surface by hybridizing in the aptamers of electrode surface with modification, wherein telluride
Cadmium/tellurium/cadmium selenide core shell quantum dot is abbreviated as CdTe@CdSe, forms ZnO NRs-NSs/CdS:The dual sensitization knots of Mn/CdTe@CdSe
Structure realizes initial photo-signal amplification；Carcinomebryonic antigen makes 5 ' ends be marked with CdTe@by specific recognition aptamers
The DNA probe solution of CdSe hybridizes and is detached from electrode surface, and CdTe@CdSe sensibilizations is caused to disappear, and reduces photo-signal,
In addition carcinomebryonic antigen has big space steric effect with the biological composite that aptamers are formed, and further decreases photoelectric current letter
Number, the Sensitive Detection to carcinomebryonic antigen is realized based on above-mentioned signal amplification mode.
In order to solve the above-mentioned technical problem, the present invention is realized by following measures：
（1）Prepare ZnO NRs-NSs/FTO electrodes；
（2）Synthesize CdS:Mn：0.5-0.8 mmol cadmium nitrates and 0.04-0.07 mmol manganese acetates are dissolved in 50 mL bis-
In secondary water, after being heated to 70 DEG C under magnetic stirring, the vulcanized sodium that a concentration of 0.03-0.06 M that 10 mL are newly prepared are added is molten
Liquid, continuation are heated to reflux 2-5 h at 70 DEG C, and the precipitation of acquisition is used to respectively washing 3 times of ethyl alcohol and secondary water respectively, then will
It is dissolved in secondary water, and the supernatant liquor for collecting yellow obtains CdS:Mn；
（3）Prepare CdTe@CdSe；
（4）It marks CdTe@CdSe on DNA probe, takes 1 mL by step（3）CdTe@CdSe of middle acquisition and a concentration of
1- (3- the dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides of 10 mM and the N- hydroxysuccinimidyls acyl of a concentration of 20 mM are sub-
The mixed liquor of amine composition is added in the phosphate buffer solution for the pH 7.4 that 1 mL contains DNA probe, and DNA probe used is dense
Degree is 10 nmol, and 5 ' ends carry amino, at 37 DEG C after 2 h of oscillating reactions, stands 12 h, is removed using microporous pipe not anti-
The label of acquisition is finally had the phosphate that the DNA probe of CdSe is dispersed in 1 mL pH 7.4 again by the DNA probe answered
In buffer solution, wherein phosphate buffer solution is abbreviated as PBS, 1- (3- dimethylamino-propyls) -3- ethyl carbodiimide hydrochlorides
Salt is abbreviated as EDC, and n-hydroxysuccinimide is abbreviated as NHS, and the DNA probe for being marked with CdTe@CdSe is abbreviated as CdTe@
CdSe-DNA；
（5）The structure of optical electro-chemistry immunosensor：By step（1）The ZnO NRs-NSs/FTO electrodes of middle acquisition impregnate
In the solution of a concentration of 2% 3- aminopropyl triethoxysilanes, hatch 1 h, after being washed with 7.4 PBS of pH, on its surface
20 μ L of drop coating are by step（2）The CdS of middle acquisition:The NHS compositions of the EDC and a concentration of 20 mM of Mn and a concentration of 10 mM mix
Liquid is closed, after hatching 2 h at room temperature, the unreacted reagent of removing wash with 7.4 PBS of pH, by a concentration of 5.0 μM of 10 μ L's
Aptamers drop coating with carcinomebryonic antigen specific recognition is to electrode surface, and aptamers 3 ' used, which are held, carries amino, and at 4 DEG C
12 h of lower hatching are washed with 7.4 PBS of pH after removing extra aptamers, continue 20 μ L of drop coating, 1 mM 6- hydroxyls -1- oneself
Mercaptan after hatching 1 h at room temperature, is washed, then by 20 μ L steps for blocking nonspecific binding site with 7.4 PBS of pH
Suddenly（4）The CdTe@CdSe-DNA drop coatings of middle acquisition hatch 2 h to electrode surface and at 37 DEG C, are washed and are removed with 7.4 PBS of pH
After removing extra CdTe@CdSe-DNA, the carcinomebryonic antigen of 10 μ L various concentrations of drop coating uses pH after hatching 2 h at 37 DEG C
7.4 PBS are washed, and complete the structure of optical electro-chemistry immunosensor；
（6）Photo-signal detects：By step（5）The ZnO NRs-NSs/FTO electrodes of middle acquisition, platinum to electrode and
Photo-signal detection is carried out in the three-electrode system of Ag/AgCl reference electrodes composition, detection method used is current-vs-time
Curve method, voltage are 0.0 V, and excitation wavelength range is 200-2500 nm, switch an excitation light source switch, institute every 10 s
Detection solution is that 5 mL contain 7.4 PBS solutions of pH of 0.01 M hydrogen peroxide, and to lead to using preceding detection solution
15 min of nitrogen deoxygenation, with the increase of carcinomebryonic antigen concentration, photo-signal gradually decreases, and passes through carcinomebryonic antigen concentration and light
The quantitative detection to carcinomebryonic antigen may be implemented in relationship between current signal.
It is of the present invention prepare ZnO NRs-NSs/FTO electrodes the specific steps are：
（1）Electrodeposition process prepares ZnO NRs/FTO electrodes：Ethyl alcohol, acetone, secondary water is used to be cleaned by ultrasonic 10 successively FTO
After min, in the zinc acetate seed solution of a concentration of 40 mM of the surfaces FTO spin coating, rotating speed is 1000 r/min, and the time is 30 s, so
Afterwards by the FTO after spin coating at 120 DEG C dry 10 min, after above-mentioned " spin coating-drying " process of repetitive operation 6 times, electric by FTO
Pole, Ag/AgCl reference electrodes and platinum carry out electro-deposition operation to the three-electrode system that electrode forms, and sedimentation potential 0.0V sinks
Product electrolyte is by the second of the ammonium acetate and a concentration of 50-80 mM of the zinc nitrate of a concentration of 30-60 mM, a concentration of 40-60 mM
The mixed liquor of diamines composition, depositing temperature are 60-80 DEG C, and sedimentation time is 12 h, and secondary water and anhydrous second are used after the completion of deposition
Alcohol washs and dry 30 min at 60 DEG C；
（2）Hydro-thermal method prepares ZnO NRs-NSs/FTO electrodes：The insertion of above-mentioned acquisition ZnO NRs/FTO electrodes is contained 10
In 25 mL autoclaves of mL mixed liquors, and make it is conductive place downwards, the mixed liquor by a concentration of 8-12 mM nitric acid
The sodium citrate of zinc, the hexamethylenetetramine of a concentration of 8-12 mM and a concentration of 0.5-2 mM forms, and 6- is heated at 60-70 DEG C
9 h, it is to be cooled to after room temperature, 30 min are calcined with secondary water washing and at 350 DEG C, finally obtain ZnO NRs-NSs/FTO
Electrode.
It is of the present invention prepare CdTe@CdSe the specific steps are：
（1）Prepare CdTe nanometer crystalline body：The sodium citrate of 0.3-0.6 mmol cadmium nitrates and 0.1-0.3 g dehydrations is dissolved in
In 50 mL secondary waters, 50-55 μ L 3- mercaptopropionic acids are then added, extremely with the sodium hydrate regulator solution pH of a concentration of 1 M
After 10.5,0.1-0.2 mmol sodium tellurites and 45-60 mg sodium borohydrides is added, 1-3 h are heated to reflux at 100 DEG C, most
It is afterwards that the mixed liquor after reaction is multiple with normal propyl alcohol centrifuge washing, and obtained dry 30 min that are deposited at 60 DEG C are obtained
CdTe nanometer crystalline body；
（2）Prepare CdTe@CdSe：Sodium hydrogen selenide solution is prepared first, by 0.2-0.3 mmol selenium powders and 0.8-1.2
Mmol sodium borohydrides are dissolved in the secondary water of 10 mL deoxygenations, under magnetic stirring, 60 DEG C of 50 min of oil bath heating, whole process
Carry out under nitrogen protection；Then the solution containing divalent cadmium ion is prepared, by 0.2-0.5 mmol cadmium nitrates and 30-35 μ
L 3- mercaptopropionic acids are dissolved in the secondary water of 10 mL deoxygenations, then extremely with the sodium hydrate regulator solution pH of a concentration of 1 M
10.5；CdTe@CdSe are finally prepared, after above-mentioned acquisition CdTe nanometer crystalline body to be dissolved in the secondary water of 25 mL deoxygenations, use is a concentration of
The sodium hydrate regulator solution pH to 10.5 of 1 M, under the protection of nitrogen, first 1-5 mL, which are slowly added dropwise, contains divalent cadmium
The solution of ion, then 1-5 mL sodium hydrogen selenide solution is slowly added dropwise, the heating reaction 20-60 min at 70-90 DEG C will
The reaction mixture of acquisition is multiple with normal propyl alcohol centrifuge washing, and finally obtained dry 30 min that are deposited at 60 DEG C lay equal stress on
Newly it is dispersed in the secondary water of 25 mL deoxygenations, it is spare.
Beneficial effects of the present invention：
（1）ZnO NRs-NSs/FTO electrodes prepared by the present invention, with good electric conductivity, biocompatibility and big
Specific surface area can greatly increase the load capacity of quantum dot and signaling molecule, signal Analysis is further amplified, to improve inspection
The sensitivity of survey.
（2）Utilize CdS:Mn and CdTe@CdSe are sensitized ZnO jointly, and the polynary sensitization structure of formation not only can be effectively
Inhibit the compound of electron-hole pair and the absorption to visible light can be enhanced, greatly enhances photo-signal, further carry
The sensitivity of high analyte.
（3）The present invention replaces recognition element of traditional antibody as carcinomebryonic antigen using aptamers, can greatly drop
Harmonic analysis operation complexity and cost, in addition, using carcinomebryonic antigen specific recognition aptamers induction DNA probe solution hybridization and
It is detached from electrode surface, the biological composite itself for causing CdTe@CdSe sensibilizations disappearances and carcinomebryonic antigen to be formed with aptamers
Space steric effect realize signal amplification, it is sensitive, efficient which amplifies mode, treatment for clinical cancer and diagnosis tool
There is important meaning.
Specific implementation mode:
For a better understanding of the present invention, with reference to the embodiment content that the present invention is furture elucidated, but the present invention
Content is not limited solely to the following examples.
Embodiment 1:Optical electro-chemistry immunosensor is used for the detection of carcinomebryonic antigen
（1）ZnO NRs-NSs/FTO electrodes are prepared, ZnO NRs/FTO electrodes are prepared first with electrodeposition process：By FTO
It is molten in the zinc acetate seed of a concentration of 40 mM of the surfaces FTO spin coating after using ethyl alcohol, acetone, secondary water to be cleaned by ultrasonic 10 min successively
Liquid, rotating speed are 1000 r/min, and the time is 30 s, then by the FTO after spin coating at 120 DEG C dry 10 min, in repetitive operation
After stating " spin coating-drying " process 6 times, in the three-electrode system being made of to electrode FTO electrodes, Ag/AgCl reference electrodes and platinum
Electro-deposition operation, sedimentation potential 0.0V are carried out, deposited electrolyte is zinc nitrate, a concentration of 50 mM by a concentration of 50 mM
Ammonium acetate and a concentration of 70 ethylenediamine composition mixed liquor, depositing temperature be 70 DEG C, sedimentation time be 12 h, deposition complete
It is washed afterwards with secondary water and absolute ethyl alcohol and dries 30 min at 60 DEG C；Then hydro-thermal method is utilized to prepare ZnO NRs-NSs/FTO
Electrode：It is inserted into ZnO NRs/FTO electrodes are obtained among the above in the 25 mL autoclaves containing 10 mL mixed liquors, and makes conducting surface
It places downwards, the mixed liquor is by the zinc nitrate of a concentration of 10 mM, the hexamethylenetetramine and a concentration of 1 of a concentration of 10 mM
The sodium citrate of mM forms, and heats 6 h at 70 DEG C, to be cooled to after room temperature, and 30 are calcined with secondary water washing and at 350 DEG C
Min finally obtains ZnO NRs-NSs/FTO electrodes；
（2）Synthesize CdS:Mn：0.6 mmol cadmium nitrates and 0.06 mmol manganese acetates are dissolved in 50 mL secondary waters,
After being heated to 70 DEG C under magnetic agitation, the sodium sulfide solution for a concentration of 0.05 M that 10 mL are newly prepared is added, continues at 70 DEG C
Under be heated to reflux 3 h, the precipitation of acquisition is used to respectively washing 3 times of ethyl alcohol and secondary water respectively, is then dissolved in secondary water, receives
The supernatant liquor for collecting yellow obtains CdS:Mn；
（3）CdTe@CdSe are prepared, prepare CdTe nanometer crystalline body first：The lemon that 0.5 mmol cadmium nitrates and 0.2 g are dehydrated
Lemon acid sodium is dissolved in 50 mL secondary waters, and 52 μ L 3- mercaptopropionic acids are then added, and is adjusted with the sodium hydroxide of a concentration of 1 M molten
After liquid pH to 10.5,0.1 mmol sodium tellurites and 50 mg sodium borohydrides are added, 2 h are heated to reflux at 100 DEG C, finally will
Mixed liquor after reaction is multiple with normal propyl alcohol centrifuge washing, and obtained dry 30 min that are deposited at 60 DEG C are obtained CdTe
Nanocrystal；Then CdTe@CdSe are prepared：Sodium hydrogen selenide solution is prepared first, by 0.25 mmol selenium powders and 1.0 mmol boron hydrogen
Change sodium to be dissolved in the secondary water of 10 mL deoxygenations, under magnetic stirring, 60 DEG C of 50 min of oil bath heating, whole process is in nitrogen
Protection is lower to be carried out；Then the solution containing divalent cadmium ion is prepared, 0.3 mmol cadmium nitrates and 32 μ L 3- mercaptopropionic acids is molten
In the secondary water of 10 mL deoxygenations, then with the sodium hydrate regulator solution pH to 10.5 of a concentration of 1 M；Finally prepare CdTe@
CdSe：After above-mentioned acquisition CdTe nanometer crystalline body to be dissolved in the secondary water of 25 mL deoxygenations, adjusted with the sodium hydroxide of a concentration of 1 M
PH value of solution is to 10.5, under the protection of nitrogen, the solution that 3 mL contain divalent cadmium ion is slowly first added dropwise, then slowly
3 mL sodium hydrogen selenide solution are added dropwise, 30 min of heating reaction at 80 DEG C will obtain the centrifugation of reaction mixture normal propyl alcohol and wash
It washs repeatedly, is finally deposited at 60 DEG C dry 30 min by what is obtained and is dispersed in again in the secondary water of 25 mL deoxygenations, it is standby
With；
（4）It marks CdTe@CdSe on DNA probe, takes 1 mL by step（3）CdTe@CdSe of middle acquisition and a concentration of
The mixed liquor of the NHS of the EDC of 10 mM and a concentration of 20 mM compositions is added in 7.4 PBS of pH that 1 mL contains DNA probe,
A concentration of 10 nmol of DNA probe used, 5 ' ends carry amino, at 37 DEG C after 2 h of oscillating reactions, stand 12 h, utilize
Microporous pipe removes unreacted DNA probe, and the CdTe@CdSe-DNA of acquisition are finally dispersed in 1 mL pH, 7.4 PBS again
In；
（5）The structure of optical electro-chemistry immunosensor：By step（1）The ZnO NRs-NSs/FTO electrodes of middle acquisition impregnate
In the solution of a concentration of 2% 3- aminopropyl triethoxysilanes, hatch 1 h, after being washed with 7.4 PBS of pH, on its surface
20 μ L of drop coating are by step（2）The CdS of middle acquisition:The NHS compositions of the EDC and a concentration of 20 mM of Mn and a concentration of 10 mM mix
Liquid is closed, after hatching 2 h at room temperature, the unreacted reagent of removing wash with 7.4 PBS of pH, by a concentration of 5.0 μM of 10 μ L's
Aptamers drop coating with carcinomebryonic antigen specific recognition is to electrode surface, and aptamers 3 ' used, which are held, carries amino, and at 4 DEG C
12 h of lower hatching are washed with 7.4 PBS of pH after removing extra aptamers, continue 20 μ L of drop coating, 1 mM 6- hydroxyls -1- oneself
Mercaptan after hatching 1 h at room temperature, is washed, then by 20 μ L steps for blocking nonspecific binding site with 7.4 PBS of pH
Suddenly（4）The CdTe@CdSe-DNA drop coatings of middle acquisition hatch 2 h to electrode surface and at 37 DEG C, are washed and are removed with 7.4 PBS of pH
After removing extra CdTe@CdSe-DNA, the carcinomebryonic antigen of 10 μ L various concentrations of drop coating uses pH after hatching 2 h at 37 DEG C
7.4 PBS are washed, and complete the structure of optical electro-chemistry immunosensor；
（6）Photo-signal detects：By step（5）The ZnO NRs-NSs/FTO electrodes of middle acquisition, platinum to electrode and
Photo-signal detection is carried out in the three-electrode system of Ag/AgCl reference electrodes composition, detection method used is current-vs-time
Curve method, voltage are 0.0 V, and excitation wavelength range is 200-2500 nm, switch an excitation light source switch, institute every 10 s
Detection solution is that 5 mL contain 7.4 PBS solutions of pH of 0.01 M hydrogen peroxide, and to lead to using preceding detection solution
15 min of nitrogen deoxygenation, with the increase of carcinomebryonic antigen concentration, photo-signal gradually decreases, and passes through carcinomebryonic antigen concentration and light
The quantitative detection to carcinomebryonic antigen may be implemented in relationship between current signal.
SEQUENCE LISTING
<110>University Of Ji'nan
<120>Detect the preparation and application of the optical electro-chemistry immunosensor of carcinomebryonic antigen
<130> 2017
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 18
<212> DNA
<213>It is artificial synthesized
<400> 1
ataccagctt attcaatt 18
<210> 2
<211> 12
<212> DNA
<213>It is artificial synthesized
<400> 2
aattgaataa gc 12
Claims (3)
1. a kind of preparation method of the optical electro-chemistry immunosensor of detection carcinomebryonic antigen, it is characterized in that including the following steps：
（1）Zinc oxide nano rod-nanometer sheet of layered structure is grown in the tin dioxide transparent conductive glass surface of doping fluorine,
The tin dioxide transparent conductive glass of middle doping fluorine is abbreviated as FTO, and zinc oxide nano rod-nanometer sheet of layered structure writes a Chinese character in simplified form ZnO
NRs-NSs prepares ZnO NRs-NSs/FTO electrodes；
（2）Synthesize the cadmiumsulfide quantum dot of additive Mn：0.5-0.8 mmol cadmium nitrates and 0.04-0.07 mmol manganese acetates is molten
In 50 mL secondary waters, after being heated to 70 DEG C under magnetic stirring, a concentration of 0.03-0.06 M that 10 mL are newly prepared are added
Sodium sulfide solution, continuation is heated to reflux 2-5 h at 70 DEG C, the precipitation of acquisition used to the respectively washing 3 of ethyl alcohol and secondary water respectively
It is secondary, it is then dissolved in secondary water, the supernatant liquor for collecting yellow obtains the cadmiumsulfide quantum dot of additive Mn, wherein additive Mn
Cadmiumsulfide quantum dot be abbreviated as CdS:Mn；
（3）Cadmium telluride/tellurium/cadmium selenide core shell quantum dot is prepared, CdTe@CdSe are abbreviated as；
（4）It marks CdTe@CdSe on DNA probe, takes 1 mL by step（3）The CdTe@CdSe of middle acquisition and a concentration of 10 mM
1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides and a concentration of 20 mM n-hydroxysuccinimide composition
Mixed liquor be added in the phosphate buffer solution for the pH 7.4 that 1 mL contains DNA probe, DNA probe used a concentration of 10
Nmol, 5 ' ends carry amino, at 37 DEG C after 2 h of oscillating reactions, stand 12 h, and unreacted DNA is removed using microporous pipe
Probe, the phosphate-buffered for finally having the DNA probe of CdSe to be dispersed in 1 mL pH 7.4 again the label of acquisition are molten
In liquid, wherein phosphate buffer solution is abbreviated as PBS, and 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides are write a Chinese character in simplified form
For EDC, n-hydroxysuccinimide is abbreviated as NHS, and the DNA probe for being marked with CdTe@CdSe is abbreviated as CdTe@CdSe-DNA；
（5）The quantitative detection of carcinomebryonic antigen：By step（1）The ZnO NRs-NSs/FTO electrodes of middle acquisition are immersed in a concentration of 2%
3- aminopropyl triethoxysilanes solution in, hatch 1 h, after being washed with 7.4 PBS of pH, 20 μ L of its surface drop coating by
Step（2）The CdS of middle acquisition:The mixed liquor of the NHS compositions of the EDC and a concentration of 20 mM of Mn and a concentration of 10 mM, in room temperature
It after 2 h of lower hatching, is washed with 7.4 PBS of pH and removes unreacted reagent, by a concentration of 5.0 μM of 10 μ L's and carcinomebryonic antigen
For the aptamers drop coating of specific recognition to electrode surface, aptamers 3 ' used end carries amino, and hatches 12 h at 4 DEG C,
It is washed with 7.4 PBS of pH after removing extra aptamers, continues 20 μ L of drop coating, 1 mM 6- hydroxyl -1- hexyl mercaptans for blocking
Nonspecific binding site after hatching 1 h at room temperature, is washed with 7.4 PBS of pH, then by 20 μ L steps（4）Middle acquisition
CdTe@CdSe-DNA drop coatings hatch 2 h to electrode surface and at 37 DEG C, CdTe@CdSe-DNA by with modification in electrode
The aptamers hybridization on surface is captured to electrode surface, is washed with 7.4 PBS of pH after removing extra CdTe@CdSe-DNA, drop
The carcinomebryonic antigen of 10 μ L various concentrations is applied, carcinomebryonic antigen is modified the aptamers in electrode surface by specific recognition, made
CdTe@CdSe-DNA solutions hybridize and are detached from electrode surface, are washed, are modified with 7.4 PBS of pH after hatching 2 h at 37 DEG C
ZnO NRs-NSs/FTO electrodes；By modifying ZnO NRs-NSs/FTO electrodes, platinum is to electrode and Ag/AgCl reference electrodes
Photo-signal detection is carried out in the three-electrode system of composition, detection method used is current versus time curve method, and voltage is
0.0 V, excitation wavelength range are 200-2500 nm, switch an excitation light source switch every 10 s, detection solution used is
5 mL contain 7.4 PBS solutions of pH of 0.01 M hydrogen peroxide, and to lead to 15 min of nitrogen deoxygenation using preceding detection solution,
With the increase of carcinomebryonic antigen concentration, photo-signal gradually decreases, by between carcinomebryonic antigen concentration and photo-signal
The quantitative detection to carcinomebryonic antigen may be implemented in relationship.
2. special according to a kind of preparation method of the optical electro-chemistry immunosensor of detection carcinomebryonic antigen described in claims 1
Sign is, it is described prepare ZnO NRs-NSs/FTO electrodes the specific steps are：
（1）Electrodeposition process prepares ZnO NRs/FTO electrodes：Ethyl alcohol, acetone, secondary water is used to be cleaned by ultrasonic 10 min successively FTO
Afterwards, in the zinc acetate seed solution of a concentration of 40 mM of the surfaces FTO spin coating, rotating speed is 1000 r/min, and the time is 30 s, then
By the FTO after spin coating at 120 DEG C dry 10 min, after above-mentioned " spin coating-drying " process of repetitive operation 6 times, by FTO electrodes,
Ag/AgCl reference electrodes and platinum carry out electro-deposition operation, sedimentation potential 0.0V, deposition electricity to the three-electrode system that electrode forms
Liquid is solved as by the ethylenediamine of the ammonium acetate and a concentration of 50-80 mM of the zinc nitrate of a concentration of 30-60 mM, a concentration of 40-60 mM
The mixed liquor of composition, depositing temperature are 60-80 DEG C, and sedimentation time is 12 h, is washed with secondary water and absolute ethyl alcohol after the completion of deposition
It washs and dries 30 min at 60 DEG C；
（2）Hydro-thermal method prepares ZnO NRs-NSs/FTO electrodes：Above-mentioned acquisition ZnO NRs/FTO electrodes are inserted into and are mixed containing 10 mL
In the 25 mL autoclaves for closing liquid, and make it is conductive place downwards, the mixed liquor by a concentration of 8-12 mM zinc nitrate, dense
The sodium citrate composition for the hexamethylenetetramine and a concentration of 0.5-2 mM that degree is 8-12 mM, 6-9 h are heated at 60-70 DEG C,
It is to be cooled to after room temperature, 30 min are calcined with secondary water washing and at 350 DEG C, finally obtain ZnO NRs-NSs/FTO electrodes.
3. special according to a kind of preparation method of the optical electro-chemistry immunosensor of detection carcinomebryonic antigen described in claims 1
Sign is, it is described prepare CdTe@CdSe the specific steps are：
（1）Prepare CdTe nanometer crystalline body：The sodium citrate of 0.3-0.6 mmol cadmium nitrates and 0.1-0.3 g dehydrations is dissolved in 50
In mL secondary waters, 50-55 μ L 3- mercaptopropionic acids are then added, with the sodium hydrate regulator solution pH to 10.5 of a concentration of 1 M
Afterwards, 0.1-0.2 mmol sodium tellurites and 45-60 mg sodium borohydrides is added, 1-3 h are heated to reflux at 100 DEG C, finally will
Mixed liquor after reaction is multiple with normal propyl alcohol centrifuge washing, and obtained dry 30 min that are deposited at 60 DEG C are obtained CdTe
Nanocrystal；
（2）Prepare CdTe@CdSe：Sodium hydrogen selenide solution is prepared first, by 0.2-0.3 mmol selenium powders and 0.8-1.2 mmol boron
Sodium hydride is dissolved in the secondary water of 10 mL deoxygenations, and under magnetic stirring, 60 DEG C of 50 min of oil bath heating, whole process is in nitrogen
It is carried out under gas shielded；Then the solution containing divalent cadmium ion is prepared, by 0.2-0.5 mmol cadmium nitrates and 30-35 μ L 3- mercaptos
Base propionic acid is dissolved in the secondary water of 10 mL deoxygenations, then with the sodium hydrate regulator solution pH to 10.5 of a concentration of 1 M；Finally
CdTe@CdSe are prepared, after above-mentioned acquisition CdTe nanometer crystalline body to be dissolved in the secondary water of 25 mL deoxygenations, with the hydrogen-oxygen of a concentration of 1 M
Change sodium and adjust pH value of solution to 10.5, under the protection of nitrogen, first 1-5 mL, which are slowly added dropwise, contains the molten of divalent cadmium ion
Liquid, then 1-5 mL sodium hydrogen selenide solution is slowly added dropwise, the heating reaction 20-60 min at 70-90 DEG C, by the anti-of acquisition
It answers mixed liquor normal propyl alcohol centrifuge washing multiple, is finally deposited at 60 DEG C dry 30 min by what is obtained and is dispersed in again
It is spare in the secondary water of 25 mL deoxygenations.
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