CN105241943B - Detect the preparation and application of the electrochemical immunosensor of circulating tumor cell - Google Patents
Detect the preparation and application of the electrochemical immunosensor of circulating tumor cell Download PDFInfo
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Abstract
The invention discloses a kind of sensor of differential pulse voltammetry detection circulating tumor cell.The preparation of the sensor, comprises the following steps：Vanadium pentoxide nanowires, gold and silver core-shell nano are prepared according to existing method；Vanadium pentoxide nanowires gold and silver core-shell nano is combined into modification aptamers；Golden nanometer particle is modified in electrode surface, connects Thiolation aptamers afterwards, specifically binds circulating tumor cell, reconnects the aptamers of vanadium pentoxide nanowires gold and silver core-shell nano modification；The electrochemical sensor combination electrochemical workstation made is detected to circulating tumor cell；Detected using electrochemical workstation.Sensor of the invention high specificity, high sensitivity is simple to operate, and test limit is low.
Description
Technical field
The present invention relates to Cell Measurement Technique field, is in particular that a kind of be catalyzed by semiconductor produces reduction electricity
Stream, alloy enhancing reduction current, so as to detect circulating tumor cell number, belongs to overdelicate electro-chemical cells immunosensor
Technology of preparing.
Background technology
In recent years, immunoassay is the clinical means of an important tumor disease in clinical examination.All the time, it is pernicious
Tumour i.e. cancer are a serious threats of human health and life security.Cancer cell, it is latent to refer to that some possess division
A kind of cell of energy caused neoformation there occurs vicious transformation and Clonal hyperplasia in the presence of carcinogenic factor.And cancer
Cell in addition to the growth of itself is out of control, can also invade arround normal structure or even can be via the internal circulatory system or leaching
Bar system is transferred to the other parts of body, so as to cause body lesion even dead.So the diagnosis of cancer early stage and early
Treatment have great importance.But it is difficult to be found that the cancer cell content of cancer early stage is relatively low, the cancer cell of low content
Measure become the important pursuit of analysis personnel.
Immunoassay, which is mainly based upon, uses antibody（Or antigen）Alternatively property chemical reagent is (or anti-to analyze measure antigen
Body) and haptens.Its proposition and development is that one of the greatest achievement of the bioanalytical chemistry estimation whole world will be carried out every year
The immunoassay of several hundred million times.Develop in immunoassay soon, using wide first-elected radio immunoassay（RIA）, RIA major advantages
It is automation equipment, easy to operate, high sensitivity, sample preparation is simple；Lived simultaneously as radiation is not present in clinical sample
Property material, thus disturb few.However, RIA needs to carry out hot operation, instrument price is again expensive, and this just excites scientists
Make great efforts the immunoassay method of exploration nonradioactive labeling, develop ELISA rapidly（ELISA）, electrochemiluminescent immunoassay
Analysis（LIA）, electrochemical immunoanalytical（ECIA）The methods of.But these immunoassay methods are typically time-consuming longer and possess
The operating process such as cumbersome sample-adding, incubation, washing, and contact of the operating personnel with immune substance is more, may be to operation
The health of personnel damages.And the detection means that some finding speeds are fast, high sensitivity, detection property are low needs instrument
It is expensive, it is impossible to be popularized in developing country.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of high sensitivity, detection speed is fast, reagent dosage is few, detection
The electrochemical immunosensor of circulating tumor cell.
In order to solve the above-mentioned technical problem, the present invention is realized by following measures：One kind detection circulating tumor is thin
The preparation of the electrochemical sensor of born of the same parents and application process, it comprises the following steps：
（1）Vanadium pentoxide nanowires, gold and silver core-shell nano are prepared according to existing method；
（2）Vanadium pentoxide nanowires-gold and silver core-shell nano is combined into modification aptamers；
（3）Golden nanometer particle is modified in electrode surface, connects Thiolation aptamers afterwards, specifically binds circulating tumor
Cell, reconnect the aptamers of vanadium pentoxide nanowires-gold and silver core-shell nano modification；
（4）The electrochemical sensor combination electrochemical workstation made is detected to circulating tumor cell.
Vanadium pentoxide nanowires of the present invention-gold and silver core-shell nano combines modification aptamers and electrochemical sensing
The preparation and application of device comprise the following steps：
（1）The preparation of vanadium pentoxide nanowires：It is characterized in that water miscible vanadic sulfate and oxidant potassium bromate are existed
Mix at room temperature, the min of magnetic stirring reaction 30, after by nitric acid reduce acidity, by mixture aqueous solution in autoclave with 180
DEG C reaction 24 h, take out autoclave be cooled to room temperature, secondary water repeatedly rinses to obtain thick yellow precipitate, will be deposited in 80 DEG C of baking ovens
In be dried for standby；
（2）The preparation of golden nanometer particle storing solution：It is characterized in that the mM of 1.25 mL 10 gold chloride is added into 42.65 mL
Secondary water, it is placed in the three neck round bottom flask of 100 mL capacity, 30 min is heated in the case where magnetic stirs, it is rear to add 2.5 mL
1 % sodium citrate, solution is first changed into pewter from yellow in 10 min, eventually becomes soft pink colour, solution is by centrifugation and two
Secondary washing, after be redispersed in secondary water, obtain golden nanometer particle storing solution；
（3）The preparation of gold and silver core-shell nano：It is characterized in that by 7 mL（2）The golden nanometer particle storing solution of preparation with
The % polyvinylpyrrolidones of 0.65 mL 0.1 mix, and magnetic stirs 20 min, the rear beta-schardinger dextrin for adding the mM of 1.5 mL 0.05 and
The mM of 1.5 mL 0.05 cysteine, magnetic stirring a few minutes mix.Add sodium hydroxide solution and hatch 10 min adjustment pH extremely
10, by 85 °C of water-baths of mixed solution.Continuous magnetic stirs 30 min afterwards, and the mM of 3 mL 0.1 silver nitrate solution, drop is added dropwise
During adding, solution is gradually changed into glassy yellow from pink colour, obtains gold and silver core-shell nano；
（4）The preparation of vanadium pentoxide nanowires-gold and silver core-shell nano：It is characterized in that will（1）Five oxidations prepared
Two vanadium nano wires take 10 mg to be dissolved in 1 mL secondary waters, add 1 % chlorination hexadiene dimethyl amine solution, ultrasound 30
Min, centrifugation washing 4 times are rear to add（3）The mL of gold and silver core-shell nano 0.5 of preparation, magnetic stir 1 h, pelleting centrifugation washing
It is repeatedly constant to color, finally obtain vanadium pentoxide nanowires-gold and silver core-shell nano；
（5）Prepare the aptamers of vanadium pentoxide nanowires-gold and silver core-shell nano modification：It is characterized in that first by 0.1
The pH to 9 of M sodium hydroxide adjustment vanadium pentoxide nanowires-gold and silver core-shell nano, rear 5 μM of mercaptan for adding 50 μ L
Change aptamers, mixed solution under the conditions of 4 °C magnetic stir 12 h, after washed with phosphate buffer solution, discard supernatant liquor,
It will be precipitated and dissolved in containing 1 mM Ca2+, 1 mM Mn2+In the incubation buffer of 0.1 % bovine serum albumin(BSA)s, 4 °C of decentralizations
Purchase use；
（6）The modification of glass-carbon electrode：It is characterized in that cleaned glass-carbon electrode is inserted to the mM of 2mL 1 gold chloride ,-
The s of electro-deposition 40 under 0.2 V voltages, golden nanometer particle in modification, phosphate buffer solution are rinsed, dried standby；
（7）Will（6）The glass-carbon electrode modified connects Thiolation aptamers, it is characterized in that being sealed by 6- sulfydryl -1- hexanols
Site is filled, then captures circulating tumor cell, the adaptation of connection vanadium pentoxide nanowires-gold and silver core-shell nano modification
Body, electrode insertion are included in 0.01 M phosphate buffer solutions of 25 μM of thionines, are passed through differential pulse voltammetry and are detected reduction electricity
Stream obtains result.
The wash solution used in the present invention is pH 7.4 phosphate buffer solution.
Circulating tumor cell of the present invention is breast cancer cell（MCF-7 and T47D）.
Beneficial effects of the present invention：
（1）The present invention is measured using differential pulse voltammetry, and operation is quick and easy, reaction and result by instrument from
It is dynamic to complete and record, the influence of subjective factor is avoided, and ensure there is good repeatability；
（2）By the use of the vanadic anhydride that bioactivity is stable, catalytic property is good as catalyst, and vanadic anhydride
Small molecule dyes can be catalyzed under the conditions of existing for hydrogen peroxide, produce reduction current；
（3）Gold and silver core-shell nano has high conductivity, while is exaggerated reduction current, plays amplified signal effect.
Brief description of the drawings
Fig. 1 show flow chart of the present invention.
A show golden nanometer particle；B show aptamers；C show circulating tumor cell；D show vanadic anhydride
Nano wire；E show gold and silver core-shell nano.
Embodiment
A kind of preparation of electrochemical sensor for detecting circulating tumor cell and application process, it comprises the following steps：
（1）Vanadium pentoxide nanowires, gold and silver core-shell nano are prepared according to existing method；
（2）Vanadium pentoxide nanowires-gold and silver core-shell nano is combined into modification aptamers；
（3）Golden nanometer particle is modified in electrode surface, connects Thiolation aptamers afterwards, specifically binds circulating tumor
Cell, reconnect the aptamers of vanadium pentoxide nanowires-gold and silver core-shell nano modification；
（4）The electrochemical sensor combination electrochemical workstation made is detected to circulating tumor cell.
Vanadium pentoxide nanowires of the present invention-gold and silver core-shell nano combines modification aptamers and electrochemical sensing
The preparation and application of device comprise the following steps：
（1）The preparation of vanadium pentoxide nanowires：It is characterized in that water miscible vanadic sulfate and oxidant potassium bromate are existed
Mix at room temperature, the min of magnetic stirring reaction 30, after by nitric acid reduce acidity, by mixture aqueous solution in autoclave with 180
DEG C reaction 24 h, take out autoclave be cooled to room temperature, secondary water repeatedly rinses to obtain thick yellow precipitate, will be deposited in 80 DEG C of baking ovens
In be dried for standby；
（2）The preparation of golden nanometer particle storing solution：It is characterized in that the mM of 1.25 mL 10 gold chloride is added into 42.65 mL
Secondary water, it is placed in the three neck round bottom flask of 100 mL capacity, 30 min is heated in the case where magnetic stirs, it is rear to add 2.5 mL
1 % sodium citrate, solution is first changed into pewter from yellow in 10 min, eventually becomes soft pink colour, solution is by centrifugation and two
Secondary washing, after be redispersed in secondary water, obtain golden nanometer particle storing solution；
（3）The preparation of gold and silver core-shell nano：It is characterized in that by 7 mL（2）The golden nanometer particle storing solution of preparation with
The % polyvinylpyrrolidones of 0.65 mL 0.1 mix, and magnetic stirs 20 min, the rear beta-schardinger dextrin for adding the mM of 1.5 mL 0.05 and
The mM of 1.5 mL 0.05 cysteine, magnetic stirring a few minutes mix.Add sodium hydroxide solution and hatch 10 min adjustment pH extremely
10, by 85 °C of water-baths of mixed solution.Continuous magnetic stirs 30 min afterwards, and the mM of 3 mL 0.1 silver nitrate solution, drop is added dropwise
During adding, solution is gradually changed into glassy yellow from pink colour, obtains gold and silver core-shell nano；
（4）The preparation of vanadium pentoxide nanowires-gold and silver core-shell nano：It is characterized in that will（1）Five oxidations prepared
Two vanadium nano wires take 10 mg to be dissolved in 1 mL secondary waters, add 1 % chlorination hexadiene dimethyl amine solution, ultrasound 30
Min, centrifugation washing 4 times are rear to add（3）The mL of gold and silver core-shell nano 0.5 of preparation, magnetic stir 1 h, pelleting centrifugation washing
It is repeatedly constant to color, finally obtain vanadium pentoxide nanowires-gold and silver core-shell nano；
（5）Prepare the aptamers of vanadium pentoxide nanowires-gold and silver core-shell nano modification：It is characterized in that first by 0.1
The pH to 9 of M sodium hydroxide adjustment vanadium pentoxide nanowires-gold and silver core-shell nano, rear 5 μM of mercaptan for adding 50 μ L
Change aptamers, mixed solution under the conditions of 4 °C magnetic stir 12 h, after washed with phosphate buffer solution, discard supernatant liquor,
It will be precipitated and dissolved in containing 1 mM Ca2+, 1 mM Mn2+In the incubation buffer of 0.1 % bovine serum albumin(BSA)s, 4 °C of decentralizations
Purchase use；
（6）The modification of glass-carbon electrode：It is characterized in that cleaned glass-carbon electrode is inserted to the mM of 2mL 1 gold chloride ,-
The s of electro-deposition 40 under 0.2 V voltages, golden nanometer particle in modification, phosphate buffer solution are rinsed, dried standby；
（7）Will（6）The glass-carbon electrode modified connects Thiolation aptamers, encapsulates site by 6- sulfydryl -1- hexanols, connects
Capture circulating tumor cell, the aptamers of connection vanadium pentoxide nanowires-gold and silver core-shell nano modification, electrode insertion
In the 0.01 M phosphate buffer solutions comprising 25 μM of thionines, reduction current is detected by differential pulse voltammetry and obtains result.
Embodiment 1（Breast cancer cell, MCF-7）
（1）The MCF-7 that selection is easily shifted with lymphatic system is measured；：
（2）The preparation of vanadium pentoxide nanowires：Water miscible vanadic sulfate and oxidant potassium bromate are mixed at room temperature
Close, the min of magnetic stirring reaction 30, after by nitric acid reduce acidity, by mixture aqueous solution in autoclave with 180 DEG C reaction 24
H, take out autoclave be cooled to room temperature, secondary water repeatedly rinses to obtain thick yellow precipitate, will be deposited in 80 DEG C of baking ovens dry it is standby
With；
（3）The preparation of golden nanometer particle storing solution：The mM of 1.25 mL 10 gold chloride adds 42.65 mL secondary waters, puts
In the three neck round bottom flask of 100 mL capacity, 30 min are heated in the case where magnetic stirs, the rear lemon for adding the % of 2.5 mL 1
Lemon acid sodium, solution is first changed into pewter from yellow in 10 min, eventually becomes soft pink colour, solution by centrifugation and secondary washing,
After be redispersed in secondary water, obtain golden nanometer particle storing solution；
（4）The preparation of gold and silver core-shell nano：By 7 mL（2）The golden nanometer particle storing solution and 0.65 mL of preparation
0.1 % polyvinylpyrrolidones mix, and magnetic stirs 20 min, the rear beta-schardinger dextrin and 1.5 mL for adding the mM of 1.5 mL 0.05
0.05 mM cysteine, magnetic stirring a few minutes mix.Add sodium hydroxide solution and hatch 10 min adjustment pH to 10, will be mixed
Close 85 °C of water-baths of solution.Continuous magnetic stirs 30 min afterwards, and the mM of 3 mL 0.1 silver nitrate solution is added dropwise, and process is added dropwise
In, solution is gradually changed into glassy yellow from pink colour, obtains gold and silver core-shell nano；
（5）The preparation of vanadium pentoxide nanowires-gold and silver core-shell nano：Will（1）The vanadic anhydride nanometer of preparation
Line takes 10 mg to be dissolved in 1 mL secondary waters, adds 1 % chlorination hexadiene dimethyl amine solution, 30 min of ultrasound, centrifugal water
Wash 4 times, it is rear to add（3）The mL of gold and silver core-shell nano 0.5 of preparation, magnetic stir 1 h, and pelleting centrifugation is washed repeatedly to color
It is constant, finally obtain vanadium pentoxide nanowires-gold and silver core-shell nano；
（6）Prepare the aptamers of vanadium pentoxide nanowires-gold and silver core-shell nano modification：First by 0.1 M hydrogen-oxygen
Change the pH to 9 of sodium adjustment vanadium pentoxide nanowires-gold and silver core-shell nano, latter add 50 μ L 5 μM Thiolation
SYL3C aptamers, mixed solution under the conditions of 4 °C magnetic stir 12 h, after washed with phosphate buffer solution, it is clear to discard upper strata
Liquid, it will be precipitated and dissolved in containing 1 mM Ca2+, 1 mM Mn2+In the incubation buffer of 0.1 % bovine serum albumin(BSA)s, under 4 °C
Place standby；
（7）The modification of glass-carbon electrode：Cleaned glass-carbon electrode insertion 2mL 1 mM gold chloride, under -0.2 V voltages
The s of electro-deposition 40, golden nanometer particle in modification, phosphate buffer solution are rinsed, dried standby；
（8）Will（7）The glass-carbon electrode modified connects Thiolation aptamers, encapsulates site by 6- sulfydryl -1- hexanols, connects
Capture MCF-7 cells, the SYL3C aptamers of connection vanadium pentoxide nanowires-gold and silver core-shell nano modification, electrode is inserted
Enter in the 0.01 M phosphate buffer solutions comprising 25 μM of thionines, reduction current is detected by differential pulse voltammetry and tied
Fruit.
Circulating tumor cell MCF-7 is detected, the range of linearity of the electrochemical immunosensor is 50 cells
mL-1~1× 107 cells·mL-1, detect and be limited to 31 cells mL-1。
Embodiment 2（Breast cancer cell, T47D）
（1）The T47D that selection is easily shifted with lymphatic system is measured；
（2）The preparation of vanadium pentoxide nanowires：Water miscible vanadic sulfate and oxidant potassium bromate are mixed at room temperature
Close, the min of magnetic stirring reaction 30, after by nitric acid reduce acidity, by mixture aqueous solution in autoclave with 180 DEG C reaction 24
H, take out autoclave be cooled to room temperature, secondary water repeatedly rinses to obtain thick yellow precipitate, will be deposited in 80 DEG C of baking ovens dry it is standby
With；
（3）The preparation of golden nanometer particle storing solution：The mM of 1.25 mL 10 gold chloride adds 42.65 mL secondary waters, puts
In the three neck round bottom flask of 100 mL capacity, 30 min are heated in the case where magnetic stirs, the rear lemon for adding the % of 2.5 mL 1
Lemon acid sodium, solution is first changed into pewter from yellow in 10 min, eventually becomes soft pink colour, solution by centrifugation and secondary washing,
After be redispersed in secondary water, obtain golden nanometer particle storing solution；
（4）The preparation of gold and silver core-shell nano：By 7 mL（2）The golden nanometer particle storing solution and 0.65 mL of preparation
0.1 % polyvinylpyrrolidones mix, and magnetic stirs 20 min, the rear beta-schardinger dextrin and 1.5 mL for adding the mM of 1.5 mL 0.05
0.05 mM cysteine, magnetic stirring a few minutes mix.Add sodium hydroxide solution and hatch 10 min adjustment pH to 10, will be mixed
Close 85 °C of water-baths of solution.Continuous magnetic stirs 30 min afterwards, and the mM of 3 mL 0.1 silver nitrate solution is added dropwise, and process is added dropwise
In, solution is gradually changed into glassy yellow from pink colour, obtains gold and silver core-shell nano；
（5）The preparation of vanadium pentoxide nanowires-gold and silver core-shell nano：Will（1）The vanadic anhydride nanometer of preparation
Line takes 10 mg to be dissolved in 1 mL secondary waters, adds 1 % chlorination hexadiene dimethyl amine solution, 30 min of ultrasound, centrifugal water
Wash 4 times, it is rear to add（3）The mL of gold and silver core-shell nano 0.5 of preparation, magnetic stir 1 h, and pelleting centrifugation is washed repeatedly to color
It is constant, finally obtain vanadium pentoxide nanowires-gold and silver core-shell nano；
（6）Prepare the aptamers of vanadium pentoxide nanowires-gold and silver core-shell nano modification：First by 0.1 M hydrogen-oxygen
Change the pH to 9 of sodium adjustment vanadium pentoxide nanowires-gold and silver core-shell nano, latter add 50 μ L 5 μM Thiolation
SYL3C aptamers, mixed solution under the conditions of 4 °C magnetic stir 12 h, after washed with phosphate buffer solution, it is clear to discard upper strata
Liquid, it will be precipitated and dissolved in containing 1 mM Ca2+, 1 mM Mn2+In the incubation buffer of 0.1 % bovine serum albumin(BSA)s, under 4 °C
Place standby；
（7）The modification of glass-carbon electrode：Cleaned glass-carbon electrode insertion 2mL 1 mM gold chloride, under -0.2 V voltages
The s of electro-deposition 40, golden nanometer particle in modification, phosphate buffer solution are rinsed, dried standby；
（8）Will（7）The glass-carbon electrode modified connects Thiolation aptamers, by 6- sulfydryl -1- hexanols（MCH）Encapsulate position
Point, then capture T47D cells, the SYL3C aptamers of connection vanadium pentoxide nanowires-gold and silver core-shell nano modification, electricity
In 0.01 M phosphate buffer solutions of the pole insertion comprising 25 μM of thionines, reduction current is detected by differential pulse voltammetry and obtained
To result.
Cancer cell T47D is detected, the range of linearity of the electrochemical immunosensor is 50 cells mL-1~1×
107 cells·mL-1, detect and be limited to 40 cells mL-1。
Claims (1)
1. a kind of preparation method for the electrochemical sensor for detecting circulating tumor cell, it comprises the following steps：
（1）Prepare vanadium pentoxide nanowires：Water miscible vanadic sulfate is mixed at room temperature with oxidant potassium bromate, magnetic
The min of stirring reaction 30, after by nitric acid reduce acidity, by mixture aqueous solution in autoclave in 180 DEG C react 24 h, take
Go out autoclave and be cooled to room temperature, secondary water repeatedly rinses to obtain thick yellow precipitate, is deposited in 80 DEG C of baking ovens and is dried for standby；
（2）Prepare golden nanometer particle storing solution：The mM of 1.25 mL 10 gold chloride is added into 42.65 mL secondary waters, is placed in 100
In the three neck round bottom flask of mL capacity, 30 min are heated in the case where magnetic stirs, the rear sodium citrate for adding the % of 2.5 mL 1,
Solution is first changed into pewter from yellow in 10 min, eventually becomes soft pink colour, solution is by centrifugation and secondary washing, rear redisperse
In secondary water, golden nanometer particle storing solution is obtained；
（3）Prepare gold and silver core-shell nano：Golden nanometer particle storing solution and the % polyethylene of 0.65 mL 0.1 prepared by 7 mL
Pyrrolidones mixing magnetic stirs 20 min, half Guang of the rear beta-schardinger dextrin and the mM of 1.5 mL 0.05 for adding the mM of 1.5 mL 0.05
Propylhomoserin, magnetic stirring a few minutes mix；Add sodium hydroxide solution and hatch 10 min adjustment pH to 10, by 85 °C of water of mixed solution
Bath；Continuous magnetic stirs 30 min afterwards, the mM of 3 mL 0.1 silver nitrate solution is added dropwise, during dropwise addition, solution is gradually by powder
Discoloration is glassy yellow, obtains gold and silver core-shell nano；
（4）Prepare vanadium pentoxide nanowires-gold and silver core-shell nano：The vanadium pentoxide nanowires of preparation are taken into 10 mg
It is dissolved in 1 mL secondary waters, adds 1 % chlorination hexadiene dimethyl amine solution, 30 min of ultrasound, centrifugation is washed 4 times, after
The mL of gold and silver core-shell nano 0.5 prepared is added, magnetic stirs 1 h, and pelleting centrifugation washing is repeatedly constant to color, finally obtains
Obtain vanadium pentoxide nanowires-gold and silver core-shell nano；
（5）Vanadium pentoxide nanowires-gold and silver core-shell nano is combined into modification aptamers：First by 0.1 M sodium hydroxide
The pH to 9 of vanadium pentoxide nanowires-gold and silver core-shell nano is adjusted, rear 5 μM of Thiolation aptamers for adding 50 μ L, is mixed
Close solution under the conditions of 4 °C magnetic stir 12 h, after washed with phosphate buffer solution, discard supernatant liquor, will be precipitated and dissolved in
Contain 1 mM Ca2+, 1 mM Mn2+In the incubation buffer of 0.1 % bovine serum albumin(BSA)s, 4 °C are transferred use of purchasing；
（6）By cleaned glass-carbon electrode insertion 2mL 1 mM gold chloride, the s of electro-deposition 40 under -0.2 V voltages, in modification
Golden nanometer particle, phosphate buffer solution are rinsed, and are dried standby, are connected Thiolation aptamers afterwards, are sealed by 6- sulfydryl -1- hexanols
Site is filled, then specifically binds circulating tumor cell, reconnects vanadium pentoxide nanowires-gold and silver core-shell nano modification
Aptamers.
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| CN106198662B (en) * | 2016-08-31 | 2018-11-09 | 济南大学 | Electrochemical method that is a kind of while detecting two kinds of tumor markers |
| CN108344783B (en) * | 2018-01-23 | 2020-04-24 | 东南大学 | Electrochemical cell sensor and preparation method and application thereof |
| CN109696465A (en) * | 2019-01-04 | 2019-04-30 | 中国药科大学 | A kind of circulating tumor cell detection method based on plasma-reinforced electrochemical reaction |
| CN109709174A (en) * | 2019-02-18 | 2019-05-03 | 武汉大学 | The enzyme type electrochemical sensor and the preparation method and application thereof of single core-shell nano line |
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| CN104459132A (en) * | 2014-12-12 | 2015-03-25 | 济南大学 | Preparation method and application of pancreatic cancer immunosensor based on gold electrodeposition and Au@Ag/CuO-GS as markers |
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| WO2014081393A1 (en) * | 2012-11-21 | 2014-05-30 | Singapore Health Services Pte Ltd | Method for detection or an antigen |
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| CN104568923A (en) * | 2014-12-01 | 2015-04-29 | 浙江省肿瘤医院 | Method and kit for detecting circulating tumor cell antigens in peripheral blood through electrochemical luminescence detection |
| CN104459132A (en) * | 2014-12-12 | 2015-03-25 | 济南大学 | Preparation method and application of pancreatic cancer immunosensor based on gold electrodeposition and Au@Ag/CuO-GS as markers |
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