CN101750489A - Beta 2-adrenergic receptor stimulant detection method based on microfluidic chip - Google Patents
Beta 2-adrenergic receptor stimulant detection method based on microfluidic chip Download PDFInfo
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- CN101750489A CN101750489A CN200810229858A CN200810229858A CN101750489A CN 101750489 A CN101750489 A CN 101750489A CN 200810229858 A CN200810229858 A CN 200810229858A CN 200810229858 A CN200810229858 A CN 200810229858A CN 101750489 A CN101750489 A CN 101750489A
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Abstract
The invention discloses a Beta 2-adrenergic receptor stimulant detection method based on microfluidic chip and is characterized in that: the detection process concretely and sequentially includes of anti-body coating, confining liquid sealing, scrubbing, competitive immunoreaction of enzyme-labeled antigen and the detected antigen and anti-body, enzymatic substrate reaction and LIF (laser induced fluorescence) detection. The invention has the advantages of short detection time, low LOD, good repeatability, easy operation, less sample dosage and integration, also has great foreseeable scientific value and economic value.
Description
Technical field
The present invention relates to chemistry and physics, be specifically related to the chip immuno analytical method; A kind of β based on micro-fluidic chip is provided especially
2-adrenoceptor agonists detection method.
Background technology
β
2-adrenoceptor agonists class medicine is a class synthetic medicine, is mainly used in control people, animal bronchial astehma and bronchial spasm.β
2-adrenoceptor agonists can be used for strengthening sportsman, animal (as horse) muscle in sports tournament, improve sports achievement, and the International Olympic Committee is with β
2-adrenoceptor agonists is classified forbidden drugs as.Studies show that the eighties both at home and abroad, adds β in feed
2-adrenoceptor agonists has nutrition reallocation effect, can obviously improve lean meat percentage.Human body has eaten and has contained β
2The food of-adrenoceptor agonists class medicine has many bad reactions.To β
2The detection of-adrenoceptor agonists is at the clinical medicine dynamic metabolism, and sports and food security aspect are all significant.
In the prior art, detect β
2The method of-adrenoceptor agonists and platform mainly contain following several: chromatographic technique, EIA enzyme immunoassay technology, gold-marking immunity detection technique and biosensor technique.β
2-adrenoceptor agonists residue detection is from taking all factors into consideration, the enzyme-linked immunosorbent assay of porous plate (belonging to the EIA enzyme immunoassay technology) is to detect the clenobuterol hydrochloride best approach at present, but incubation time on the orifice plate and immune response required time are longer, complete operation that can not robotization.Though chromatography is finally to confirm analytical approach, it is loaded down with trivial details and expense is high, shortcomings such as instrument costliness to have an operation.Gold-marking immunity detection technique and biosensor technique are confined to single part of mensuration of single, can't realize high flux at present.
People catch at a kind of technique effect better based on the β of micro-fluidic chip
2-adrenoceptor agonists detection system and based on the detection method of this system.
Summary of the invention
The purpose of this invention is to provide a kind of β based on micro-fluidic chip
2-adrenoceptor agonists detection method.
The contrast prior art, micro-fluidic chip has high flux, and robotization is integrated, and is consuming time few, fast, advantage such as efficient, so we combine micro-fluidic chip and realize β with enzyme-linked immunoassay method
2The fast detecting of-adrenoceptor agonists.
β based on micro-fluidic chip of the present invention
2-adrenoceptor agonists detection method is concrete according to the β based on micro-fluidic chip
2-adrenoceptor agonists detection system is operated,
The content of described detection system now is described below:
Described detection system is for using the detection system of laser detection; It is characterized in that:
Described detection system is a core with micro-fluidic chip 1, and described micro-fluidic chip 1 constitutes by three-decker is compound successively: the fluid passage layer 101 on upper strata, middle elastic film layer 102, the pneumatic control layer 103 of lower floor; This three part physical relationship each other is as follows: fluid passage layer 101 and pneumatic key-course 103 by its separately with elastic film layer 102 between surface tension closely link to each other.
Include 2～100 groups of detecting units 201 in the described detection system, detecting unit 201 specifically includes following ingredient: little valve cell 202, analysis channel 203, sample liquid pool 204;
Also be connected with on described each detecting unit 201 and supply with buffer pool 205 and the waste liquid pool 206 that at least one detecting unit 201 uses.
Described detection system preferably realizes sample introduction by the little valve cell 202 that is arranged on the micro-fluidic chip 1: described little valve cell 202 specifically is to be one group of use more than 3 or 3, and it moves by programmed control; Being constructed as follows of described little valve cell 202: elastic film layer 102, be arranged on dam structure 207 on the fluid passage layer 101, be arranged on the gas cavity 208 that is in the corresponding position on the pneumatic control layer 103 of elastic film layer 102 opposite sides with dam structure 207.
Described detection system specifically includes following ingredient: micro-fluidic chip 1, object lens 2, spectroscope 3, condenser lens 4, pin hole 5, emission light optical filter 6, photomultiplier 7, signal picker 8, exciting light optical filter 9 laser instruments 10; Wherein:, arranging object lens 2, spectroscope 3, condenser lens 4, pin hole 5, emission light optical filter 6, photomultiplier 7 successively according to straight line substantially according to distance micro-fluidic chip 1 order from the close-by examples to those far off; Signal picker 8 is connecting photomultiplier 7, a side of spectroscope 3 according to from the close-by examples to those far off be disposed in order exciting light optical filter 9, laser instrument 10.
In the described detection system, in testing process, it is 470～650nm that effective operation wavelength of laser instrument 10 requires; Detect under the laser excitation condition of the laser instrument 10 that satisfies this requirement, thing to be detected sends the fluorescence of 560～620nm.
For the ease of operation, in the described detection system, each detecting unit 201 structure is identical or become the mirror image symmetry, and the analysis channel 203 of each detecting unit 201 is concentrated at surveyed area and arranged;
The width of the gas channels 209 in described analysis channel 203 and/or the pneumatic control layer 103 and the degree of depth are in 10～300 microns scope.
In described micro-fluidic chip with three-decker, be furnished with gas channels 209 in the described pneumatic control layer 103, each gas channels 209 is being controlled the gas cavity 208 of 2～100 groups of arranged in series, and described gas cavity 208 correspondences little valve cell 202 correspondingly;
Gas channels 209 by pilot-gas cavity 208 switching gas and carry the size of tolerance and then control little valve cell 202.
Described β based on micro-fluidic chip
2In-adrenoceptor agonists the detection system, fluid passage layer 101 and pneumatic key-course 103 employed materials can be glass in the micro-fluidic chip 1, and elastic film layer 102 materials can be the PDMS polymkeric substance.
In the described detection system, any 3 little valve cells 202 specifically are the little valve cells 202 of adjacent three of arranging on same gas channels 209 not;
In the described detection system, possess the functipnal capability that detects a plurality of detecting units 201 simultaneously, specific practice is: the same position straight line in 50 second time at each split tunnel moves the laser instrument acquired signal.
The present invention is based on the β based on micro-fluidic chip of said detecting system
2-adrenoceptor agonists detection method specifically is described below:
β based on micro-fluidic chip
2-adrenoceptor agonists detection method, its described testing process specifically is successively: antibody sandwich, confining liquid sealing, washing, enzyme-labelled antigen and determined antigen and antibody competition immune response, substrate for enzymatic activity reaction and laser-Induced Fluorescence Detection.
Described β based on micro-fluidic chip
2In-adrenoceptor agonists the detection method, use elastic film layer 102 as the immunoreactive solid phase adsorption carrier of competition, determined antigen and enzyme-labelled antigen are as the immune response that is at war with of competition thing and the lip-deep antibody of PDMS.
Described β
2In-adrenoceptor agonists the detection method, realize sample introduction by the little valve cell 202 that is arranged on the micro-fluidic chip 1: described little valve cell 202 specifically is to be one group of use more than 3 or 3, and it moves by programmed control; Being constructed as follows of described little valve cell 202: elastic film layer 102, be arranged on dam structure 207 on the fluid passage layer 101, be arranged on the gas cavity 208 that is in the corresponding position on the pneumatic control layer 103 of elastic film layer 102 opposite sides with dam structure 207; The sample introduction operating process moves by programmed control; When elastic deformation takes place elastic film layer 102, produce negative pressure by micro-fluidic chip 1, under the effect of negative pressure, the flow direction waste liquid pool 206 in sample liquid pool 204, the buffer pool 205.
Described β
2In-adrenoceptor agonists the detection method, employed elastic film layer 102 specifically is the PDMS film.It has hydrophobicity, has the characteristic of strong adhesion protein class material.
Described β
2In-adrenoceptor agonists the detection method, use the non-blooming substrate A DHP (10-Acetyl-3 of horseradish peroxidase enzyme catalytic, 7-dihydroxyphenoxazine, 10-acetyl group-3,7-dihydroxy phenoxazine) generates the product resorufin (resorufin, the different phenoxazine oxazolone of 9-hydroxyl-3-) that fluorescence is arranged;
During concrete the detection, require to detect under wavelength is the laser excitation condition of 470～650nm, thing to be detected can send the fluorescence of 560～620nm.
Described β
2In-adrenoceptor agonists the detection method, detect a plurality of passages simultaneously; Specific practice is: the same position straight line in 50 second time at each analysis channel 203 moves the laser instrument acquired signal.
The invention provides a kind of β
2The method for quick of-adrenoceptor agonists and micro-fluidic chip special.The present invention is based on the competitive enzyme-linked immune adsorption reaction, and laser-induced fluorescence (LIF) is a detection means, compares with traditional multiaperture-type enzyme linked immunological adsorption reaction, and reagent consumption is low, and sensitivity is greatly improved.
Among the present invention, the material of fluid passage layer 101 and pneumatic key-course 103 is a glass, and the material of middle elastic film layer 2 is the PDMS polymkeric substance.Bilevel glass can reuse, and the PDMS film is removable.
Gas channels 209 in the described chip pneumatic control layer 103 exports 210 places in gas circuit and is connected the input of solenoid control gases at high pressure or negative-pressure gas by external pipeline with solenoid valve.The switching of the gas of different pressures causes the vibration of elastic film layer 102.During the gases at high pressure input, elastic film layer 102 is close to the dam structure 207 of fluid passage layer 101, and little valve cell 202 is closed, and fluid is static; During the negative-pressure gas input, middle elastic film layer 102 is bent downwardly, and little valve cell 202 is opened, and fluid flows.The wriggling of any three continuous little valve cells 202 can produce the effect of Micropump, the travelling forward of propelling fluid.β on the whole integrated chip
2The testing process of-adrenoceptor agonists is controlled automatically with the little valve of pneumatic Micropump.
The micro-fluidic chip technology is the research focus of current instrumental analysis, this technology is mainly based on analytical chemistry and biological chemistry, utilize micro electronmechanical process technology, process the microchannel network of 10-300 micron in silicon, glass, quartz, Polymer Surface, be driving force mainly with electroosmotic flow and electrophoresis stream, by changing driving voltage, flow direction and the speed of control fluid in the network of microchannel, not only be easy to realize operation such as sampling, dilution, enrichment, extraction, mixing, reaction, separation, detection, and be easy to the operating unit array to target analytes.Up to now, on the chip platform that is integrated with pneumatic micro valve unit 202 to β
2-adrenoceptor agonists carries out the method for the parallel detection of many groups and does not appear in the newspapers.The present invention has creatively designed and has been used for β
2The array micro-fluidic chip that-adrenoceptor agonists detects detects when can realize a plurality of sample.Whole process was finished in 30 minutes, and sample consumption is at micro updating.The present invention compares with traditional microwell plate method, advantage such as has and amount of samples is few direct, quick, simple to operate.
In a word, the present invention can finish the β in a plurality of samples on glass-PDMS hybridization hybrid chip of more than one square centimeters at short notice
2Detect in the time of-adrenoceptor agonists.In terms of existing technologies, the present invention have directly, control fast and automatically, simple to operate and characteristics that amount of samples is few.It has foreseeable huge scientific value and economic worth.
Description of drawings
The present invention is further detailed explanation below in conjunction with drawings and the embodiments:
Fig. 1 is the β based on micro-fluidic chip
2-adrenoceptor agonists detection system theory of constitution synoptic diagram;
Fig. 2 is that the structure of micro-fluidic chip 1 is formed principle schematic, contains eight detecting units 201;
Fig. 3 is that the integral body of micro-fluidic chip 1 is formed the synoptic diagram vertical view;
Fig. 4 is fluid passage layer 101 structural representation;
Fig. 5 is pneumatic control layer 103 structural representation;
Fig. 6 is structure 207 position views in dam in the liquid road;
Fig. 7 is detecting unit 201 structural principle synoptic diagram;
Fig. 8 is sectional view and three little valve cell 202 continuous motion synoptic diagram of valve arrangement;
Fig. 9 is the reappearance with variable concentrations clenobuterol hydrochloride standard items on the chip piece; Fig. 9 is β on the integrated microfluidic chip among the embodiment 1
2The repeated collection of illustrative plates that-adrenoceptor agonists detects, detected β
2-adrenoceptor agonists is clenobuterol hydrochloride (clenbuterol);
Figure 10 is the clenobuterol hydrochloride working curve;
Figure 11 is a clenobuterol hydrochloride semilog working curve;
Figure 12 is a recovery spectrogram 1;
Figure 13 is a recovery spectrogram 2; Figure 13 is the β in the pig urine sample on the integrated microfluidic chip among the embodiment 2
2The collection of illustrative plates of-adrenoceptor agonists the recovery.
Embodiment
Use a kind of β described in instructions based on micro-fluidic chip
2-adrenoceptor agonists detection system: described detection system is a core with micro-fluidic chip 1, and described micro-fluidic chip 1 constitutes by three-decker is compound successively: the fluid passage layer 101 on upper strata, middle elastic film layer 102, the pneumatic control layer 103 of lower floor; This three part physical relationship each other is as follows: fluid passage layer 101 and pneumatic key-course 103 by its separately with elastic film layer 102 between surface tension closely link to each other.
Include 2～100 groups of detecting units 201 in the described detection system, detecting unit 201 specifically includes following ingredient: little valve cell 202, analysis channel 203, sample liquid pool 204;
Also be connected with on described each detecting unit 201 and supply with buffer pool 205 and the waste liquid pool 206 that at least one detecting unit 201 uses.
Described detection system preferably realizes sample introduction by the little valve cell 202 that is arranged on the micro-fluidic chip 1: described little valve cell 202 specifically is to be one group of use more than 3 or 3, and it moves by programmed control; Being constructed as follows of described little valve cell 202: elastic film layer 102, be arranged on dam structure 207 on the fluid passage layer 101, be arranged on the gas cavity 208 that is in the corresponding position on the pneumatic control layer 103 of elastic film layer 102 opposite sides with dam structure 207.
Described detection system specifically includes following ingredient: micro-fluidic chip 1, object lens 2, spectroscope 3, condenser lens 4, pin hole 5, emission light optical filter 6, photomultiplier 7, signal picker 8, exciting light optical filter 9 laser instruments 10; Wherein:, arranging object lens 2, spectroscope 3, condenser lens 4, pin hole 5, emission light optical filter 6, photomultiplier 7 successively according to straight line substantially according to distance micro-fluidic chip 1 order from the close-by examples to those far off; Signal picker 8 is connecting photomultiplier 7, a side of spectroscope 3 according to from the close-by examples to those far off be disposed in order exciting light optical filter 9, laser instrument 10.
In the described detection system, in testing process, it is 470～650nm that effective operation wavelength of laser instrument 10 requires; Detect under the laser excitation condition of the laser instrument 10 that satisfies this requirement, thing to be detected sends the fluorescence of 560～620nm.
For the ease of operation, in the described detection system, each detecting unit 201 structure is identical or become the mirror image symmetry, and the analysis channel 203 of each detecting unit 201 is concentrated at surveyed area and arranged;
The width of the gas channels 209 in described analysis channel 203 and/or the pneumatic control layer 103 and the degree of depth are in 10～300 microns scope.
In described micro-fluidic chip with three-decker, be furnished with gas channels 209 in the described pneumatic control layer 103, each gas channels 209 is being controlled the gas cavity 208 of 2～100 groups of arranged in series, and described gas cavity 208 correspondences little valve cell 202 correspondingly; Gas channels 209 by pilot-gas cavity 208 switching gas and carry the size of tolerance and then control little valve cell 202.
Described β based on micro-fluidic chip
2In-adrenoceptor agonists the detection system, fluid passage layer 101 and pneumatic key-course 103 employed materials can be glass in the micro-fluidic chip 1, and elastic film layer 102 materials can be the PDMS polymkeric substance.
In the described detection system, any 3 little valve cells 202 specifically are the little valve cells 202 of adjacent three of arranging on same gas channels 209 not;
In the described detection system, possess the functipnal capability that detects a plurality of detecting units 201 simultaneously, specific practice is: the same position straight line in 50 second time at each split tunnel moves the laser instrument acquired signal.
β based on said detecting system based on micro-fluidic chip
2-adrenoceptor agonists detection method is described below:
β based on micro-fluidic chip
2-adrenoceptor agonists detection method, its described testing process specifically is successively: antibody sandwich, confining liquid sealing, washing, enzyme-labelled antigen and determined antigen and antibody competition immune response, substrate for enzymatic activity reaction and laser-Induced Fluorescence Detection.
Described β based on micro-fluidic chip
2In-adrenoceptor agonists the detection method, use elastic film layer 102 as the immunoreactive solid phase adsorption carrier of competition, determined antigen and enzyme-labelled antigen are as the immune response that is at war with of competition thing and the lip-deep antibody of PDMS.
Described β
2In-adrenoceptor agonists the detection method, realize sample introduction by the little valve cell 202 that is arranged on the micro-fluidic chip 1: described little valve cell 202 specifically is to be one group of use more than 3 or 3, and it moves by programmed control; Being constructed as follows of described little valve cell 202: elastic film layer 102, be arranged on dam structure 207 on the fluid passage layer 101, be arranged on the gas cavity 208 that is in the corresponding position on the pneumatic control layer 103 of elastic film layer 102 opposite sides with dam structure 207; The sample introduction operating process moves by programmed control; When elastic deformation takes place elastic film layer 102, produce negative pressure by micro-fluidic chip 1, under the effect of negative pressure, the flow direction waste liquid pool 206 in sample liquid pool 204, the buffer pool 205.
Described β
2In-adrenoceptor agonists the detection method, employed elastic film layer 102 specifically is the PDMS film.It has hydrophobicity, has the characteristic of strong adhesion protein class material.
Described β
2In-adrenoceptor agonists the detection method, use the non-blooming substrate A DHP (10-Acetyl-3 of horseradish peroxidase enzyme catalytic, 7-dihydroxyphenoxazine, 10-acetyl group-3,7-dihydroxy phenoxazine) generates the product resorufin (resorufin, the different phenoxazine oxazolone of 9-hydroxyl-3-) that fluorescence is arranged;
During concrete the detection, require to detect under wavelength is the laser excitation condition of 470～650nm, thing to be detected can send the fluorescence of 560～620nm.
Described β
2In-adrenoceptor agonists the detection method, detect a plurality of passages simultaneously; Specific practice is: the same position straight line in 50 second time at each split tunnel moves the laser instrument acquired signal.
The material of described fluid passage layer 101 and pneumatic key-course 103 is a glass, and the material of middle elastic film layer 102 is the PDMS polymkeric substance.Bilevel glass can reuse, and the PDMS film is removable.
Gas channels 209 in the described chip pneumatic control layer 103 is connected the input of solenoid control gases at high pressure or negative-pressure gas exporting 210 places in gas circuit with solenoid valve by external pipeline.The switching of the gas of different pressures causes the vibration of elastic film layer 102.During the gases at high pressure input, elastic film layer 102 is close to the dam structure 207 of fluid passage layer 101, and little valve cell 202 is closed, and fluid is static; During the negative-pressure gas input, middle elastic film layer 102 is bent downwardly, and little valve cell 202 is opened, and fluid flows.The wriggling of any three continuous little valve cells 202 can propelling fluid travel forward, can produce the effect of Micropump.β on the whole integrated chip
2The testing process of-adrenoceptor agonists is controlled automatically with the little valve of pneumatic Micropump.
Used integrated microfluidic chip designs voluntarily for this laboratory, and by the sandwich method preparation, configuration as shown in Figure 2.The fluid passage layer 101 on upper strata and the pneumatic control layer of lower floor 103 are glass sheet, adopt standard photoetching, wet etching to make.Top fluid passage layers channel size is 300 μ m * 50 μ m, and the channel size of lower floor's pneumatic control layer is 160 μ m * 70 μ m.Middle elastic film layer 102 thickness are 300 μ m, are used for forming the Micropump/little valve that transports various immunoreagents.In said chip, bilevel glass can reuse, and middle elastic film is removable, greatly reduces cost and the fabrication cycle of producing this chip like this.
On integrated chip the clenobuterol hydrochloride in the standard model being commonly called as clenbuterol hydrochloride, is typical β
2-adrenoceptor agonists class material carries out fast detecting.Antibody, sealer, the potpourri of enzyme mark clenobuterol hydrochloride and clenobuterol hydrochloride pumps into analysis channel 203 from sample liquid pool 204, and cleansing solution and substrate pump into analysis channel 203 from buffering liquid pool 205.As the immunoreactive solid phase adsorption carrier of chip, EIA enzyme immunoassay is at war with in passage with the PDMS film in this part work.Whole analytic processes is as follows: antibody sandwich → washing → confining liquid sealing → washing → enzyme mark clenobuterol hydrochloride and clenobuterol hydrochloride competition immune response → washing → substrate for enzymatic activity → laser-Induced Fluorescence Detection.Said process all carries out in the passage of micro-fluidic chip, and is finished by the little valve control of pneumatic Micropump.Antibody sandwich → washing → this part process of confining liquid sealing → washing is carried out stand-by on chip in advance.Only need enzyme mark clenobuterol hydrochloride and these four processes of clenobuterol hydrochloride competition immune response → washing → substrate for enzymatic activity → laser-Induced Fluorescence Detection during formal the detection, detection can be finished within 30 minutes.Antibody, sealer, the potpourri of enzyme mark clenobuterol hydrochloride and clenobuterol hydrochloride pumps into analysis channel 203 from sample liquid pool 204, and cleansing solution and substrate pump into analysis channel (203) from buffering liquid pool 205.
Carrying out the clenobuterol hydrochloride repeatability of 0ng/mL～10.0ng/mL concentration according to the method described above respectively measures, obtain six repetitions of variable concentrations clenobuterol hydrochloride standard items on chip piece, its relative standard deviation (RSD) is all less than 8%, and the repeated spectrogram that obtains as shown in Figure 9.Through calculating, the detectability of the described method of this patent is lower than 0.1ng/mL, and this detectability than the micro-pore plate type detection method of routine is much lower.
On integrated chip, the clenobuterol hydrochloride in the pig urine sample is carried out fast detecting.Antibody, sealer, enzyme mark clenobuterol hydrochloride pumps into analysis channel 203 with the potpourri that has added the pig urine sample of clenobuterol hydrochloride from sample liquid pool 204, and cleansing solution and substrate pump into analysis channel 203 from buffering liquid pool 205.This part work equally with the PDMS film as the immunoreactive solid phase adsorption carrier of chip, EIA enzyme immunoassay is at war with in passage.Whole analytic processes is as follows: antibody sandwich → washing → confining liquid sealing → washing → enzyme mark clenobuterol hydrochloride and clenobuterol hydrochloride competition immune response → washing → substrate for enzymatic activity → laser-Induced Fluorescence Detection.Said process all carries out in the passage of micro-fluidic chip, and is finished by the little valve control of pneumatic Micropump.Antibody sandwich → washing → this part process of confining liquid sealing → washing is carried out stand-by on chip in advance.Only need enzyme mark clenobuterol hydrochloride and these four processes of clenobuterol hydrochloride competition immune response → washing → substrate for enzymatic activity → laser-Induced Fluorescence Detection during formal the detection, detection can be finished within 30 minutes.
In containing the pig urine sample of finite concentration clenobuterol hydrochloride, add 0.5ng/mL respectively according to the method described above, 1.0ng/mL, 1.2ng/mL, the 1.5ng/mL clenobuterol hydrochloride, the recovery value that records as calculated is between 97%～124%.Clenobuterol hydrochloride recovery collection of illustrative plates as shown in Figure 12 and Figure 13 in the pig urine sample that obtains.
Claims (6)
1. based on the β of micro-fluidic chip
2-adrenoceptor agonists detection method is characterized in that:
Described testing process specifically is successively: antibody sandwich, confining liquid sealing, washing, enzyme-labelled antigen and determined antigen and antibody competition immune response, substrate for enzymatic activity reaction and laser-Induced Fluorescence Detection.
2. according to the described β of claim 1 based on micro-fluidic chip
2-adrenoceptor agonists detection method is characterized in that: described β based on micro-fluidic chip
2In-adrenoceptor agonists the detection method, use elastic film layer (102) as the immunoreactive solid phase adsorption carrier of competition, determined antigen and enzyme-labelled antigen are as the immune response that is at war with of competition thing and the lip-deep antibody of PDMS.
3. according to the described β of claim 2 based on micro-fluidic chip
2-adrenoceptor agonists detection method is characterized in that: described β
2In-adrenoceptor agonists the detection method, realize sample introduction by pump valve: described pump valve is by 3 or 3 pumps that above valve is formed, and it moves by programmed control; When elastic deformation takes place elastic film layer (102), produce negative pressure by micro-fluidic chip (1), under the effect of negative pressure, the flow direction waste liquid pool (206) in sample liquid pool (204), the buffer pool (205).
4. according to the described β of claim 3 based on micro-fluidic chip
2-adrenoceptor agonists detection method is characterized in that: described β
2In-adrenoceptor agonists the detection method, employed elastic film layer (102) specifically is the PDMS film.It has hydrophobicity, has the characteristic of strong adhesion protein class material.
5. according to one of them described β of claim 1～4 based on micro-fluidic chip
2-adrenoceptor agonists detection method is characterized in that: described β
2In-adrenoceptor agonists the detection method, use the non-blooming substrate 10-of horseradish peroxidase enzyme catalytic acetyl group-3,7-dihydroxy phenoxazine generates the product resorufin that fluorescence is arranged: the different phenoxazine oxazolone of 9-hydroxyl-3-;
During concrete the detection, require to detect under wavelength is the laser excitation condition of 470～650nm, thing to be detected can send the fluorescence of 560～620nm.
6. according to the described β of claim 5 based on micro-fluidic chip
2-adrenoceptor agonists detection method is characterized in that: described β
2In-adrenoceptor agonists the detection method, detect a plurality of passages simultaneously; Specific practice is: the same position straight line in 50 second time at each split tunnel moves the laser instrument acquired signal.
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| CN102861545A (en) * | 2012-09-13 | 2013-01-09 | 中国科学院大连化学物理研究所 | Ultraviolet photocatalytic micro-reaction chip system based on titanium dioxide nano fibers |
| CN103861668A (en) * | 2012-12-17 | 2014-06-18 | 中国科学院沈阳自动化研究所 | Microfluidic chip based on pneumatic micro pumps and micro mixer |
| CN103969240A (en) * | 2014-05-20 | 2014-08-06 | 大连理工大学 | High-flux paper micro-fluidic chip for detecting clenbuterol-like matters on basis of SERS |
| CN113275044A (en) * | 2020-02-20 | 2021-08-20 | 北京京东方健康科技有限公司 | Detection chip, use method thereof and detection device |
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| CN102861545A (en) * | 2012-09-13 | 2013-01-09 | 中国科学院大连化学物理研究所 | Ultraviolet photocatalytic micro-reaction chip system based on titanium dioxide nano fibers |
| CN102861545B (en) * | 2012-09-13 | 2015-02-04 | 中国科学院大连化学物理研究所 | Ultraviolet photocatalytic micro-reaction chip system based on titanium dioxide nano fibers |
| CN103861668A (en) * | 2012-12-17 | 2014-06-18 | 中国科学院沈阳自动化研究所 | Microfluidic chip based on pneumatic micro pumps and micro mixer |
| CN103861668B (en) * | 2012-12-17 | 2015-05-13 | 中国科学院沈阳自动化研究所 | Microfluidic chip based on pneumatic micro pumps and micro mixer |
| CN103969240A (en) * | 2014-05-20 | 2014-08-06 | 大连理工大学 | High-flux paper micro-fluidic chip for detecting clenbuterol-like matters on basis of SERS |
| CN113275044A (en) * | 2020-02-20 | 2021-08-20 | 北京京东方健康科技有限公司 | Detection chip, use method thereof and detection device |
| WO2021164530A1 (en) * | 2020-02-20 | 2021-08-26 | 京东方科技集团股份有限公司 | Detection chip and usage method therefor, and detection apparatus |
| CN113275044B (en) * | 2020-02-20 | 2022-07-12 | 北京京东方健康科技有限公司 | Detection chip, use method thereof and detection device |
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