CN108802145A - A kind of electrochemica biological sensor and preparation method thereof of detection alpha-fetoprotein - Google Patents
A kind of electrochemica biological sensor and preparation method thereof of detection alpha-fetoprotein Download PDFInfo
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- CN108802145A CN108802145A CN201810560601.6A CN201810560601A CN108802145A CN 108802145 A CN108802145 A CN 108802145A CN 201810560601 A CN201810560601 A CN 201810560601A CN 108802145 A CN108802145 A CN 108802145A
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- 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
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- 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
- G01N27/308—Electrodes, e.g. test electrodes; Half-cells at least partially made of carbon
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- 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
- G01N27/3277—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction being a redox reaction, e.g. detection by cyclic voltammetry
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- 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
- G01N27/416—Systems
- G01N27/48—Systems using polarography, i.e. measuring changes in current under a slowly-varying voltage
Abstract
The invention discloses a kind of electrochemica biological sensor of detection alpha-fetoprotein, the preparation method of the sensor includes the following steps：(1) preparation of Au@Ag core-shell nanos；(2) alpha-fetoprotein aptamers modify Au@Ag core-shell nanos；(3) p-aminobenzene sulfonic acid modification bare glassy carbon electrode is prepared；(4) electrochemica biological sensor is prepared.The present invention is prepared for a kind of electrochemica biological sensor of detection alpha-fetoprotein using with the chemically active Au@Ag core-shell nanos design of adjustable electric for the first time, and the sensor operations are convenient and have high sensitivity.
Description
Technical field
The present invention relates to biosensor technology fields, are modified based on Au@Ag core-shell nanos more particularly, to one kind
Electrode is used to detect the biosensor of alpha-fetoprotein.
Background technology
Alpha-fetoprotein is a kind of primary serum albumen of embryonic development early stage, and adult is generated by liver cell, and content is extremely low.Blood
Level of Alpha Fetoprotein is related to the outer tumour of liver cancer, embryonal tumors and part liver in clear, table of the alpha-fetoprotein in tumour cell
Up to having the characteristics that high efficiency and specificity, the severity of content and the state of an illness is closely related.Therefore Serum Alpha Fetoprotein
Content can be used as one of the early diagnosis index of primary carcinoma of liver and teratoma, also can be used as observation of curative effect and judges more afterwards
Index.
The detection method of alpha-fetoprotein mainly has fluorescence analysis, chemiluminometry, enzyme-linked immunization, chromatography at present
Analytic approach, enzyme mark electrophoresis and radioimmunology etc..However most of technologies reported still haves that operation is complicated, and price is held high
It is expensive, it the problems such as can not detect in real time.Therefore, the analysis method for developing rapid sensitive goes quantitative detection alpha-fetoprotein to its phase
Close tumour early detection, early diagnosis, early treatment, observation of curative effect and more afterwards monitoring be of great significance.Electrochemical student
Object sensor has many advantages, such as low detection limit, high sensitivity, easy to operate, of low cost.
Invention content
In view of the above-mentioned problems existing in the prior art, the present patent application provides a kind of electrochemical student of detection alpha-fetoprotein
Object sensor and preparation method thereof.The present invention is designed using with the chemically active Au@Ag core-shell nanos of adjustable electric for the first time
It is prepared for a kind of electrochemica biological sensor of detection alpha-fetoprotein, the sensor operations are convenient and have high sensitivity.
Technical scheme is as follows：
A kind of electrochemica biological sensor of detection alpha-fetoprotein, the preparation method of the sensor include the following steps：
(1) preparation of Au@Ag core-shell nanos；
(2) alpha-fetoprotein aptamers modify Au@Ag core-shell nanos；
(3) p-aminobenzene sulfonic acid modification bare glassy carbon electrode is prepared；
(4) electrochemica biological sensor is prepared.
The preparation method of the Au@Ag core-shell nanos is：At room temperature, solution of gold nanoparticles, phosphate are delayed
Fliud flushing, polyvinylpyrrolidonesolution solution mix in proportion with ascorbic acid solution, add silver nitrate solution later, are protected from light
3h, centrifugation washing, is made Au Ag core-shell nanos, isochoric formation Au Ag core-shell nano aqueous solutions are spare later.Institute
The grain size for stating Gold nanoparticle is 17.8 ± 1.8nm；The particle concentration of the solution of gold nanoparticles is 1 × 10-8mol/L；Institute
The pH for stating phosphate buffer is 7.4；The mass concentration of the polyvinylpyrrolidonesolution solution is 0.5~5%, and volumetric usage is
400~4000 μ L；The molar concentration of the ascorbic acid solution is 0.05~0.2mol/L, and volumetric usage is 500~2000 μ L；
The molar concentration of the silver nitrate solution is 1~5mmol/L, and volumetric usage is 0.1~10mL.
The method of alpha-fetoprotein aptamers modification Au@Ag core-shell nanos is：By Au@Ag made from step (1)
Core-shell nanoparticle solution is adapted to liquid solution with alpha-fetoprotein and is added in tbe buffer liquid simultaneously, and 12h is stood after vibrating mixing, it
Constant volume is spare again after centrifugation, water washing collection precipitation afterwards.The Au@Cu2A concentration of the 1 × 10 of O core-shell nanoparticle solutions-8~10
×10-8Mol/L, volumetric usage are 25~250 μ L；A concentration of 0.1~5 μm of ol/L of the carcinomebryonic antigen adaptation liquid solution, body
Product dosage is 1~20 μ L；A concentration of 0.05~0.5mol/L of the tbe buffer liquid, volumetric usage are 0.1~1mL；It is described from
Heart rotating speed is 6500r/min.
The method of glass-carbon electrode for preparing p-aminobenzene sulfonic acid modification is：Use alundum (Al2O3) polishing powder to naked first
Glass-carbon electrode is processed by shot blasting, is cleaned by ultrasonic 1 minute with ethyl alcohol and ultra-pure water, and clean rear nitrogen drying is spare；Then dry
P-aminobenzene sulfonic acid is electroplated in net glassy carbon electrode surface；Finally electrode is immersed in phosphorus pentachloride solution and activates 30 minutes, later
Taking-up cleans up spare.The grain size of the alundum (Al2O3) polishing powder is 1.0,0.5 or 0.3 μm；The plating p-aminophenyl
The system of sulfonic acid is three-electrode system, and reference electrode is saturated calomel electrode, is platinum electrode to electrode；The plating is to ammonia
The method of base benzene sulfonic acid is cyclic voltammetry, and scanning range is -0.5~0.5V, sweep speed 100mV/s, sweep time
It is 30 minutes；The molar concentration of the p-aminobenzene sulfonic acid solution is 5~50mmol/L, and volumetric usage is 1~4mL；Described five
The molar concentration of chlorination phosphorus solution is 10~100mmol/L, and volumetric usage is 1~4mL.
The method for preparing electrochemica biological sensor is：The glass of p-aminobenzene sulfonic acid modification described in step (3)
With the single stranded DNA solution of alpha-fetoprotein aptamers complementation in carbon electrodes drop coating, used after reacting 30~180 minutes at room temperature
The Au@Ag core-shell nanoparticle solutions that the aptamers made from electrode surface drop coating step (2) are modified again after water is rinsed well, together
Sample is rinsed with water spare after extra reactant after reacting 20~120 minutes；Finally using the glass-carbon electrode modified as work
Electrode, the alpha-fetoprotein solution of various concentration in electrode surface drop coating, according to the electrification of the Au@Ag core-shell nanos of gained
It learns oxidation current value and the logarithm of alpha-fetoprotein concentration is in a linear relationship, drawing curve.Described and alpha-fetoprotein aptamers
A concentration of 2~10 μm of ol/L of complementary single stranded DNA solution, volumetric usage are 2~10 μ L；The Au@Ag of the aptamers modification
A concentration of the 5 × 10 of core-shell nanoparticle solution-9~1 × 10-8Mol/L, volumetric usage are 2~10 μ L；The detection first tire
The system of albumen is three-electrode system, and reference electrode is Ag/AgCl electrodes, is platinum electrode to electrode；The detection first tire
The method of albumen is differential pulse voltammetry, and scanning range is -0.2~0.6V；Sweep speed is 4mV/s；The difference arteries and veins
The electrolyte solution for rushing voltammetry is phosphate buffer；The pH of the phosphate buffer is 7.4；The phosphate buffer
Volume be 2mL.
The present invention is beneficial to be had technical effect that：
The present invention can be regulated and controled by the shell thickness to Au@Ag core-shell nanos come its electrochemistry of system adjustment
The power of response signal；
The present invention can be steadily quantitative by Au@Ag core-shell nanos by the base pair complementarity effect between DNA chain
Electrode surface is modified, this makes it possible to obtain stable and accurate electrochemical signals；
The alpha-fetoprotein aptamers on Au@Ag core-shell nanos of the present invention surface have alpha-fetoprotein specific affine
Property, thus the electrochemica biological sensor to the identification of alpha-fetoprotein quick and precisely, there is specificity well.
Description of the drawings
Fig. 1 is the transmission electron microscope picture that Au@Ag core-shell nanos are made in the embodiment of the present invention 1.
Fig. 2 is electrochemica biological sensor made from the embodiment of the present invention 1 to the working curve of alpha-fetoprotein determination.
Specific implementation mode
With reference to the accompanying drawings and examples, the present invention is specifically described.
Embodiment 1
A kind of electrochemica biological sensor of detection alpha-fetoprotein, the preparation method of the sensor include the following steps：
(1) preparation of Au@Ag core-shell nanos：It is the gold nanoparticle of 17.8 ± 1.8nm to take grain size, and 20mL is taken to exist
10 times of concentration after ten minutes is centrifuged under the conditions of 8000r/min to be dissolved in again in 2mL ultra-pure waters.Take 4mL, the phosphate of pH=7.4 slow
Fliud flushing and the polyvinylpyrrolidonesolution solution of 2mL a concentration of 1% (wt%) are uniformly mixed, and 2mL gold nanos are added thereto later
The ascorbic acid solution of particle solution and a concentration of 0.1mol/L of 1mL, is eventually adding the silver nitrate of a concentration of 3mmol/L of 6.4mL
Solution obtains Au Ag core-shell nanos after reacting 3 hours under uniform oscillating condition, constant volume is that 1mL is spare after centrifugation washing.
The transmission electron microscope picture of gained Au@Ag core-shell nanos is as shown in Figure 1, the Au@Ag core-shell nano grains synthesized as seen from Figure 1
It is 48.7 ± 3.4nm that son, which has apparent nucleocapsid, grain size, and corresponding shell thickness is about 15.5 ± 2.6nm.
(2) alpha-fetoprotein aptamers modify Au@Ag core-shell nanos：To the tbe buffer of 100 a concentration of 0.5mol/L of μ L
The alpha-fetoprotein adaptation of Au@Ag core-shell nanoparticle solutions and 20 a concentration of 200nmol/L of μ L that 50 μ L are prepared is added in liquid
Liquid solution stands 12 hours after vibrating mixing, later by using ultra-pure water constant volume spare for 50 μ L again after centrifuging washing.
(3) p-aminobenzene sulfonic acid modifies bare glassy carbon electrode：First by bare glassy carbon electrode respectively with 1.0,0.5 and 0.3 μm
Alundum (Al2O3) polishing powder polishing treatment is dried up after being cleaned by ultrasonic respectively 1 minute with ethyl alcohol and ultra-pure water with nitrogen.It then will place
The bare glassy carbon electrode with smooth mirror surface managed is used as working electrode, and saturated calomel electrode is used as reference electrode, platinum electrode
As to electrode, cyclic voltammetry curve scanning, scanning range are carried out in the p-aminobenzene sulfonic acid solution of a concentration of 20mmol/L
For -0.5~0.5V, sweep speed 100mV/s, p-aminophenyl has totally been plated with ultrapure water after scanning 30 minutes
The glass-carbon electrode of sulfonic acid is finally dipped in after being activated 30 minutes in the phosphorus pentachloride solution of a concentration of 40mmol/L of 2mL and takes out
It cleans up spare.
(4) structure of electrochemica biological sensor：The glass carbon electricity of p-aminobenzene sulfonic acid modification first described in step (3)
The single stranded DNA solution of 5 μ L a concentration of 10 μm of ol/L and alpha-fetoprotein aptamers complementation in pole surface drop coating, after reaction 2 hours
The Au@Ag core-shell nano grains that aptamers of the 5 μ L as described in step (2) are modified in electrode surface drop coating again after being rinsed with water totally
Sub- solution, same reaction are rinsed with water spare after extra reactant after 2 hours.Then 5 μ in glassy carbon electrode surface drop coating
With the clean electrode of ultrapure water after the alpha-fetoprotein solution reaction 2h of L various concentrations, using the glass-carbon electrode modified as work
Electrode, Ag/AgCl electrodes be reference electrode, platinum electrode be to electrode scan differential pulse volt-ampere curve, scanning range be-
0.2~0.6V；Sweep speed is 4mV/s, the phosphate buffer of electrolyte solution 2mL, pH=7.4.According to the Au@of gained
The electrochemical oxidation current value and the logarithm of alpha-fetoprotein concentration of Ag core-shell nanos are in a linear relationship, drawing curve.
Working curve is as shown in Fig. 2, the detection range of this method is 1~100pg/mL as shown in Figure 2.
(5) practical application of electrochemica biological sensor：The accuracy of the electrochemical sensor is by measuring Wuxi the 4th
The concentration of hospital's liver cancer patient alpha-fetoprotein in blood is verified：First by the blood of 2mL liver cancer patients with the rotating speed of 400g from
The heart obtains 1mL serum after five minutes, and it is spare to be diluted to 2mL with PBS solution；It is obtained by chemical illumination immunity analysis instrument measurement
The concentration of middle AFP, is diluted to 20pg/mL, and the dilute serum sample that 5 μ L are prepared is taken to be added drop-wise to Au@Ag core-shell nano grains
On the electrode of son modification, Electrochemical Detection, 3 groups of parallel determination are carried out with reference to step (4), the content for measuring wherein alpha-fetoprotein is
20.6pg/mL, error 3%, testing result is more accurate.
Embodiment 2
With reference to 1 step of embodiment, a kind of electrochemica biological sensor of detection alpha-fetoprotein is prepared, clinic is applied to
Experiment：
The blood of 2mL liver cancer patients is obtained into 1mL serum after five minutes with the rotating speed centrifugation of 400g first, it is dilute with PBS solution
It releases spare to 2mL；Then the wherein actual concentrations of AFP are obtained by chemical illumination immunity analysis instrument measurement, is diluted to
45pg/mL takes the dilute serum sample that 5 μ L are prepared to be added drop-wise on the electrode of Au@Ag core-shell nanos modification, with reference to step
(4) carry out Electrochemical Detection, 3 groups of parallel determination, measure wherein alpha-fetoprotein content be 44.2pg/mL, error 1.8%,
Testing result is more accurate.
Embodiment 3
With reference to 1 step of embodiment, a kind of electrochemica biological sensor of detection alpha-fetoprotein is prepared, clinic is applied to
Experiment：
The blood of 2mL liver cancer patients is obtained into 1mL serum after five minutes with the rotating speed centrifugation of 400g first, it is dilute with PBS solution
It releases spare to 2mL.Then the wherein actual concentrations of AFP are obtained by chemical illumination immunity analysis instrument measurement, is diluted to
75pg/mL takes the dilute serum sample that 5 μ L are prepared to be added drop-wise on the electrode of Au@Ag core-shell nanos modification, with reference to step
(4) carry out Electrochemical Detection, 3 groups of parallel determination, measure wherein alpha-fetoprotein content be 72.8pg/mL, error 2.9%,
Testing result is more accurate.
Claims (9)
1. a kind of electrochemica biological sensor of detection alpha-fetoprotein, which is characterized in that the preparation method of the sensor includes
Following steps：
(1) preparation of Au@Ag core-shell nanos；
(2) alpha-fetoprotein aptamers modify Au@Ag core-shell nanos；
(3) p-aminobenzene sulfonic acid modification bare glassy carbon electrode is prepared；
(4) electrochemica biological sensor is prepared.
2. electrochemica biological sensor according to claim 1, which is characterized in that the Au@Ag core-shell nanos
Preparation method is：At room temperature, by solution of gold nanoparticles, phosphate buffer, polyvinylpyrrolidonesolution solution and Vitamin C
Acid solution mixes in proportion, adds silver nitrate solution later, is protected from light 3h, and Au Ag core-shell nano grains are made in centrifugation washing
Son, isochoric formation Au@Ag core-shell nano aqueous solutions are spare later.
3. according to the method described in claim 2, it is characterized in that, the grain size of the Gold nanoparticle is 17.8 ± 1.8nm；Institute
The particle concentration for stating solution of gold nanoparticles is 1 × 10-8mol/L；The pH of the phosphate buffer is 7.4；The polyethylene
The mass concentration of pyrrolidone solution is 0.5~5%, and volumetric usage is 400~4000 μ L；Mole of the ascorbic acid solution
A concentration of 0.05~0.2mol/L, volumetric usage are 500~2000 μ L；The molar concentration of the silver nitrate solution be 1~
5mmol/L, volumetric usage are 0.1~10mL.
4. electrochemica biological sensor according to claim 1, which is characterized in that the alpha-fetoprotein aptamers modification
The method of Au@Ag core-shell nanos is：Au@Ag core-shell nanoparticle solutions made from step (1) are adapted to alpha-fetoprotein
Liquid solution is added in tbe buffer liquid simultaneously, and 12h is stood after vibrating mixing, and constant volume is standby again after centrifugation later, water washing collection precipitation
With.
5. electrochemica biological sensor according to claim 4, which is characterized in that the Au@Cu2O core-shell nanos are molten
A concentration of the 1 × 10 of liquid-8~10 × 10-8Mol/L, volumetric usage are 25~250 μ L；The carcinomebryonic antigen adaptation liquid solution
A concentration of 0.1~5 μm of ol/L, volumetric usage are 1~20 μ L；A concentration of 0.05~0.5mol/L of the tbe buffer liquid, volume
Dosage is 0.1~1mL；The centrifugal rotational speed is 6500r/min.
6. electrochemica biological sensor according to claim 1, which is characterized in that described to prepare p-aminobenzene sulfonic acid modification
The method of glass-carbon electrode be：Bare glassy carbon electrode is processed by shot blasting with alundum (Al2O3) polishing powder first, with ethyl alcohol and is surpassed
Pure water is cleaned by ultrasonic 1 minute, and clean rear nitrogen drying is spare；Then p-aminophenyl sulphur is electroplated in clean glassy carbon electrode surface
Acid；Finally electrode is immersed in phosphorus pentachloride solution and is activated 30 minutes, taking-up later cleans up spare.
7. electrochemica biological sensor according to claim 6, which is characterized in that the grain of the alundum (Al2O3) polishing powder
Diameter is 1.0,0.5 or 0.3 μm；The system of the plating p-aminobenzene sulfonic acid is three-electrode system, and reference electrode is that saturation is sweet
Mercury electrode is platinum electrode to electrode；It is described plating p-aminobenzene sulfonic acid method be cyclic voltammetry, scanning range be-
0.5~0.5V, sweep speed 100mV/s, sweep time are 30 minutes；The molar concentration of the p-aminobenzene sulfonic acid solution is
5~50mmol/L, volumetric usage are 1~4mL；The molar concentration of the phosphorus pentachloride solution is 10~100mmol/L, and volume is used
Amount is 1~4mL.
8. electrochemica biological sensor according to claim 1, which is characterized in that described to prepare electrochemica biological sensor
Method be：Described in step (3) p-aminobenzene sulfonic acid modification glassy carbon electrode surface drop coating on alpha-fetoprotein aptamers
Complementary single stranded DNA solution, again in electrode surface drop coating step after being rinsed with water after reacting 30~180 minutes at room temperature totally
(2) the Au@Ag core-shell nanoparticle solutions that aptamers made from are modified are rinsed with water more after equally reacting 20~120 minutes
It is spare after remaining reactant；Finally using the glass-carbon electrode modified as working electrode, the various concentration in electrode surface drop coating
Alpha-fetoprotein solution, according to pair of the electrochemical oxidation current value and alpha-fetoprotein concentration of the Au@Ag core-shell nanos of gained
In a linear relationship, the drawing curve of number.
9. electrochemica biological sensor according to claim 8, which is characterized in that described complementary with alpha-fetoprotein aptamers
Single stranded DNA solution a concentration of 2~10 μm of ol/L, volumetric usage be 2~10 μ L；The Au@Ag nucleocapsids of the aptamers modification
A concentration of the 5 × 10 of nano-particle solution-9~1 × 10-8Mol/L, volumetric usage are 2~10 μ L；The detection alpha-fetoprotein
System be three-electrode system, reference electrode be Ag/AgCl electrodes, to electrode be platinum electrode；The detection alpha-fetoprotein
Method be differential pulse voltammetry, scanning range be -0.2~0.6V；Sweep speed is 4mV/s；The differential pulse volt
The electrolyte solution of peace method is phosphate buffer；The pH of the phosphate buffer is 7.4；The body of the phosphate buffer
Product is 2mL.
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Cited By (2)
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| CN110632149A (en) * | 2019-09-27 | 2019-12-31 | 昆山迪安医学检验实验室有限公司 | Electrochemical sensor for alpha fetoprotein detection and preparation method and application thereof |
| CN114755277A (en) * | 2022-03-04 | 2022-07-15 | 上海健康医学院 | Biosensor and preparation method and application thereof |
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| CN105628931A (en) * | 2016-01-19 | 2016-06-01 | 济南大学 | Preparation and application of alpha fetoprotein electrochemical immunosensor based on silver deposition |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110632149A (en) * | 2019-09-27 | 2019-12-31 | 昆山迪安医学检验实验室有限公司 | Electrochemical sensor for alpha fetoprotein detection and preparation method and application thereof |
| CN110632149B (en) * | 2019-09-27 | 2022-05-20 | 昆山迪安医学检验实验室有限公司 | Electrochemical sensor for alpha fetoprotein detection and preparation method and application thereof |
| CN114755277A (en) * | 2022-03-04 | 2022-07-15 | 上海健康医学院 | Biosensor and preparation method and application thereof |
| CN114755277B (en) * | 2022-03-04 | 2024-04-05 | 上海健康医学院 | Biosensor and preparation method and application thereof |
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