CN203630139U - Chemiluminescence immunoassay biosensor detection device - Google Patents
Chemiluminescence immunoassay biosensor detection device Download PDFInfo
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- CN203630139U CN203630139U CN201320692169.9U CN201320692169U CN203630139U CN 203630139 U CN203630139 U CN 203630139U CN 201320692169 U CN201320692169 U CN 201320692169U CN 203630139 U CN203630139 U CN 203630139U
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Abstract
The utility model discloses a chemiluminescence immunoassay biosensor detection device. The device comprises an acoustic energy transmission and chemiluminescence immunoassay reaction detection device part and a control detection part. The acoustic energy transmission and chemiluminescence immunoassay reaction detection device part comprises an ultrasonic transducer, ultrasonic coupling colloid, an acoustic focusing lens integrated glass substrate, a replaceable chemiluminescence immunoassay biosensing substrate, condensing transparent glass, a diaphragm and a light lens, and an optical signal detection circuit. The control detection part comprises an ultrasonic sensor, a proportion integration differentiation (PID) ultrasonic energy mode control module, a weak optical signal processing module, a data analysis and feedback control module and an energy control mode and chemiluminescence immunoassay reaction system database. According to the chemiluminescence immunoassay biosensor detection device, the reaction efficiency is improved, and the detection sensitivity, accuracy, repeatability and stability are expanded.
Description
Technical field
The utility model belongs to biosensor technology field, relates to a kind of chemiluminescence immunoassay biology sensor pick-up unit, particularly according to sonochemistry principle, chemiluminescence immunoassay biology sensor pick-up unit based on micro-flowing injection.
Background technology
As a kind of immune analysis method, immunity biosensor is because its selectivity is good, and analysis speed is fast, operate simple and easy, testing tool have the features such as high performance price ratio just a large amount of be applied in the every field such as medical treatment & health, food security, environmental monitoring.Under this background, how to develop performance brilliance, otherness is little, and the biology sensor that consistance is high is is still researched and developed the key topic in field always.In recent decades, immuno analytical method combines the characteristic such as sensitive and convenient of the technology such as specific recognition reaction between antibody-antigen and galvanochemistry, spectroscopy, surface acoustic wave, for example high selectivity and the high sensitivity to target analytes such as tumor markers, heavy metal ion, organic toxicants, becomes one of important analysis means of the every field such as clinical, biological chemistry, environmental analysis.And for the analyzing detecting methods such as radio immunoassay, enzyme-linked immunosorbent assay, fluoroimmunoassay, the advantages such as chemiluminescence detection has without radiocontamination, required instrument is simple, detectability is low, the dynamics range of sensitivity height and width, can combine and carry out high-quality detection analysis with multiple different sensing detection pattern neatly.
Micro-flowing injection analytical approach have convenient and flexible operation, analysis speed fast, be easy to robotization and accuracy advantages of higher.Loaded down with trivial details with respect to some conventional method of analysis operation stepss, analysis time is longer, sample consumption is large, the problems such as cost of determination height, micro-flowing injection technology combines with immunoassay and chemical luminescence detection method and the micro-flowing injection immuno analytical method that grows up has been proved to be a kind of strong determination method, and corresponding analysis and detecting instrument device has also been widely used in numerous key areas of environmental monitoring, Pharmaceutical Analysis, Food Inspection and clinical analysis.But because conventional immune response is limited to its strict demand to reaction conditions and reaction system, and affect the efficiency of reacting, caused analysis time and analytical performance still to become a major issue can not be ignored.How effectively to solve this major issue, namely in shortening analysis time as far as possible, guarantee and improve the major issue of analytical performance, become key character of the present utility model.And for the feature of number of chemical electrochemiluminescent immunoassay biological respinse system, consider from the angle of entire system, the observing and controlling parameter of chemical effect of ultrasound is combined and forms exercisable Quality Control procedural information with chemiluminescence immunoassay performance of biosensor parameter technology is also the important key feature that the utlity model has broad applicability.Currently in enzyme linked immunoassay process, there are ultrasonic and two kinds of accelerated reaction methods of frequency electromagnetic waves, comparatively speaking, ultrasonic radiation energy combines with solid-phase immunity biosensor technique and can improve when detecting analytical performance, can also keep the miniaturization of pick-up unit, for pick-up unit is widely used in the technical foundation that field monitoring facilitates.
In antibody (antigen) on immunity biosensor solid phase carrier and the zymoprotein of catalytic reaction and detected liquid, between corresponding antigen (antibody), have two alternate energy barrier---Nernst layers, how overcoming this barrier restriction raising reaction efficiency is an important problem.Even if crossed over collision or combination in the antibody/antigen generation effecting reaction yardstick of this energy barrier, yet exist the probability of certain non-specific binding, this phenomenon can be brought ground unrest and interference, causes the reduction of the performances such as detectability, sensitivity and repetition consistance.Therefore how to promote that non-specific binding changes to specific binding, and improve the efficiency changing, also will be one of major issue how improving immune response efficiency.
According to the principle of sonochemistry, in chemical reaction, ultrasonic energy brings two basic effects, the one, oscillation effect, the 2nd, energy dispersal effect.Find according to analysis to existing problem in immune response, can utilize compositions of these effect Promote immunities reactions of ultrasonic energy to pass through not the Nernst layer between co-phasal surface and effectively react.The effect of this effect is also by many experimental works are confirmed.In addition, be far longer than nonspecific adhesion for the specific binding power between antibody/antigen, ultrasonic effect also can accelerate to realize the transformation of non-specific binding to specific binding, namely promotes to arrange in conjunction with the reselecting property of albumen configuration.Reduce within the specific limits the impact of energy perturbation on specific binding by controlling ultrasonic energy intensity, ensure the conversion rates of its effective specific binding.The effect of these effects promotes reaction rate by physics mode to a certain extent, increases specific immune response efficiency and in the level of system, improves the performance such as detection lower bound, sensitivity and repetition consistance that chemiluminescence immunoassay bio-sensing detects.
Summary of the invention
The surface effect of immunity biosensor solid phase can exist important impact to immune response efficiency, and in immune sensing testing process, the diffusion restriction between antibody and antigen or other enzyme labeling things is also one of them important influence factor simultaneously.They not only affect detection lower bound and the sensitivity that in immune response, should embody, and also affect immune response speed, thereby have limited the detection efficiency of chemiluminescence immunoassay biological respinse simultaneously.The utility model is for this major issue, for these problems of chemiluminescence immunoassay biological sensing and detecting system, introduce controlled ultrasonic energy radiation function according to the principle of sonochemistry, by the energy perturbation of sensor solid phase surface region appropriateness is improved to immunoreactive speed in immunity biosensor sensitive membrane, obtain the effect of homogeneous reaction, promote degree of depth immune response, improve sensitivity and consistent repeatability that chemiluminescence immunoassay bio-sensing detects.In addition, the quality control in immune bio-sensing testing process is also a major issue can not be ignored, and the controlled ultrasonic energy radiotechnology of introducing according to the principle of sonochemistry can become the basic technology content of Quality Control process.By combining closely with Quality Control process, can standard ground obtain necessary observing and controlling parameter and performance parameter and form relation information database.This information database is the key components that form whole controlled ultrasonic energy radiochemoluminescence immunity biosensor pick-up unit system.
Feature of the present utility model is from entire system level, ultrasonic energy radiation function and chemiluminescence immunoassay biology sensor detection analysis are combined, and in the level of entire system, in conjunction with micro-flowing injection technology, the final analytical approach of measuring according to sonochemistry principle, chemiluminescence immunoassay biology sensor pick-up unit based on micro-fluidic injection and chemiluminescence immunoassay biological respinse that forms, improves the performance index such as detection lower bound, sensitivity and repetition consistance that detect analytic system.And for the feature of number of chemical electrochemiluminescent immunoassay biological respinse system, in entire system level, the observing and controlling parameter of chemical effect of ultrasound is combined with the performance parameter of chemiluminescence immunoassay biosensor technology, and form exercisable Quality Control procedural information needing in order to application widely.These are important key features of the present utility model.
The purpose of this utility model is for the deficiencies in the prior art, and a kind of chemiluminescence immunoassay biology sensor pick-up unit is provided.
The utility model device comprises acoustic energy transmission and chemiluminescence immunoassay reaction checking device part, control test section.
Described acoustic energy transmission comprises transducer fixing base 111, acoustic energy damping vibration attenuation sheet 110, ultrasonic transducer 101, ultrasonic coupling colloid 102, the integrated glass substrate 103 of sound focusing lens arra, replaceable chemiluminescence immunoassay bio-sensing substrate 104, silicone rubber O-ring 109, optically focused clear glass 107, diaphragm and optical lens 108, optical signal detecting circuit 106 from bottom to up successively with chemiluminescence immunoassay reaction checking device part.
Set gradually from bottom to up and form flow injection reaction tank/injection device by the integrated glass substrate 103 of sound focusing lens arra, silicone rubber O-ring 109, optically focused clear glass 107;
High-frequency transducer 101 is placed between macromolecule ultrasonic coupling colloid 102 and acoustic energy damping vibration attenuation sheet 110, controls the radiation intensity of high-frequency transducer's 101 ultrasonic energies by PID ultrasonic energy mode control module 202.
The integrated glass substrate 103 of sound focusing lens arra has the import and export of two passages as Fluid Transport, and two passages are located at respectively the both sides of ultrasonic coupling colloid 102; Integrated glass substrate 103 upper surfaces of sound focusing lens arra have groove, place successively from bottom to up sonac 201, replaceable chemiluminescence immunoassay bio-sensing substrate 104.This groove and two passages form inverted U-shaped structure, make fluid flow to another passage from a passage.
Be provided with silicone rubber O-ring 109 at the integrated glass substrate of sound focusing lens arra 103 edges, for seal flow injection reaction pond/injection device, regulate the height of the flowing reactive chamber 105 of flow injection reaction tank/injection device, and form enclosed construction with optically focused clear glass 107.
The height of the flowing reactive chamber 105 of described flow injection reaction tank/injection device is 1～3mm.
Described replaceable chemiluminescence immunoassay bio-sensing substrate 104 is to form the crosslinked basement membrane of carrier surface by the crosslinked glutaraldehyde of silylating reagent, shitosan etc., form basement membrane, further immobilized antigen molecule (or antibody molecule) and catalyzing enzyme on basement membrane, wherein catalyzing enzyme reacts in order to catalytic oxidation-reduction, produce electroactive material and cause curent change, the kind of conventional enzyme is alkaline phosphatase, horseradish peroxidase etc.
Described optically focused clear glass 107 is greater than unorganic glass or the macromolecule glass of 90 ﹪ for transmittance; Be carved with lensing groove at the lower surface of optically focused clear glass 107, be used for assembling chemiluminescent intensity.By adjusting the degree of depth and the radius of circle size of lensing groove, control sonac 201 sends light signal and makes focus on diaphragm and optical lens 108 surfaces through lensing groove.
In the flowing reactive chamber 105 of flow injection reaction tank/injection device, be provided with replaceable chemiluminescence immunoassay bio-sensing substrate 104, the lower surface of replaceable chemiluminescence immunoassay bio-sensing substrate 104 is stained with sonac 201, and the material of its sonac 201 is Kynoar PVDF.
Replaceable chemiluminescence immunoassay bio-sensing substrate 104 sees through successively optically focused clear glass 107 by fluorescence signal and passes to optical signal detecting circuit 106 with diaphragm and optical lens 108, and photovoltaic reaction occurs, the picking up of settling signal.
Described control test section comprises sonac 201, PID ultrasonic energy mode control module 202, faint optical signal processing module 203, data analysis and feedback control module 204, energy-controlled mode and chemiluminescence immunoassay reaction system database 205.
PID ultrasonic energy mode control module 202 is as controlled processing unit, receive the ultrasonic energy signal of sonac 201 and the feedback signal of energy-controlled mode and chemiluminescence immunoassay reaction system database 205, PID ultrasonic energy mode control module 202 drives ultrasonic transducer 101 to start mode of operation; Faint optical signal processing module 203 receives the collection signal of optically focused clear glass 107, energy-controlled mode and chemiluminescence immunoassay reaction system database 205 are by the signal of data analysis and feedback control module 204 analyzing and processing faint optical signal processing modules 203, then the signal after identification is transmitted to PID ultrasonic energy mode control module 202, finally regulate the ultrasound emission pattern (frequency and intensity) of ultrasonic transducer 101.
Utilize above-mentioned device to carry out the analytical approach of chemiluminescence immunoassay biological respinse mensuration, specifically:
Step (1). regulate the field of radiational energy of ultrasonic transducer 101
Concave spherical surface size by sound lens on ultrasonic coupling colloid 102 is focused, and making focus is 1～2mm to the vertical range on replaceable chemiluminescence immunoassay bio-sensing substrate 104 surfaces.
On described ultrasonic coupling colloid 102, the characteristic dimension of the concave spherical surface of sound lens is determined by design parameters such as the characteristic dimensions of the flowing reactive chamber 105 of energy size, macromolecule gold film electrode substrate 107 materials and thickness and the flow injection reaction tank/injection device of ultrasonic frequency, focusing.
Overall principle is: 1. regulate ultrasonic transducer 101 intensity to make the interior solution to be measured in flowing reactive chamber 105 of flow injection reaction tank/injection device can produce the disturbance of machinery and temperature, promote diffusional effect, the energy barrier that overcomes reaction, has improved reaction efficiency; 2. be adjusted to suitable intensity, can promote that in solution to be measured, non-specific binding molecular separation between antibody/antigen is reset, and change to specific binding, make solution reaction to be measured abundant.
Step (2). carry out micro-flowing injection analysis
The flowing reactive chamber 105 that solution to be measured, current-carrying are injected into flow injection reaction tank/injection device through a passage constant speed reaches the mixing of the two, finally flows out through another passage.
Carrying out in micro-flowing injection analytic process, if be fixed with antigen molecule on replaceable chemiluminescence immunoassay bio-sensing substrate 104, make the corresponding antibodies generation specific binding reaction in solution to be measured.The microminiaturization of reaction tank has been realized in the flowing reactive chamber 105 of described flow injection reaction tank/injection device by photoetching or etching method.
Step (3). ultrasound emission energy intelligent monitoring process
Carrying out in the process of step (2), the utility model carries out ultrasound emission energy intelligent monitoring simultaneously.Described Based Intelligent Control process comprises two feedback signal path, is respectively feedback signal path, the detection feedback network of ultrasonic energy.
The feedback signal of described ultrasonic energy is detected the intensity of ultrasonic radiation energy by sonac 201, itself and ultrasonic transducer 101 and PID ultrasonic energy mode control module 202 form a closed loop measurement and control system.
PID ultrasonic energy mode control module 202 receives the ultrasonic energy signal of sonac 201, and PID ultrasonic energy mode control module 202 drives ultrasonic transducer 101 to start mode of operation.
Described detection feedback network is to process detection module 203, data analysis and feedback control module 204, energy-controlled mode and chemiluminescence immunoassay reaction system database 205 by PID ultrasonic energy mode control module 202, ultra-weak electronic signal to form closed loop measurement and control system.
Ultra-weak electronic signal is processed detection module 203 and is processed the electrochemical signals that optical signal detecting circuit 106 obtains, regulate the emittance of ultrasonic transducer 101 to determine preferably immune response effect by ultrasonic energy mode control module 202, data analysis and feedback control module 204 will be integrated the parameter sets of different immune response systems, forming energy control model and chemiluminescence immunoassay reaction system database 205, the content of these databases will provide necessary pid control parameter for the application of different immune detection analysis systems.Further, utilize the measured ultrasonic energy feedback parameter of standard Quality Control immune response solution and sonac 201, the dynamic effect of the control parameter that obtains ultrasonic radiation energy to electro-chemistry immunity reaction, by dynamics data between reaction effect parameter and ultrasonic energy control parameter is carried out to matching, obtain the Optimal Parameters for controlling, these Optimal Parameters and pid control parameter carry out information fusion, obtain the concentration of target antibody in solution to be measured or antigen.
Described intelligent monitoring method, according to the feature kinetic curve of the standard immunoassay chemical reaction intensity obtaining in advance, obtains the control parameter of ultrasonic radiation energy, controls chemo-immunity reaction.
The beneficial effects of the utility model are:
The utility model device and application process utilize micro-cavitation that the principle of sonochemistry and ultrasonic energy produce to stir the technology of effectiveness binding specificity immune response and micro-flowing injection chemiluminescence analysis, develop greatly the typical micro-flowing injection chemiluminescence immunoassay technology of crosslinked glutaraldehyde (shitosan) the basement membrane immobilized antigen of silanization, and prepared immune biological sensing and detecting system.The more existing typical technology method of the method has the following advantages:
Accelerate response sample and real reaction process, greatly shortened the reaction time, improved detection efficiency, be very suitable for the on-line quick detection application in multiple fields such as clinical, environmental monitoring, food security.
A) superior consistent reappearance.Sample and micro-stirring and reaction energy transmission effects that each stage of reacting of sensitive materials produces near reaction surface because ultrasonic energy focuses on, make the homogenising of reaction interface obtain larger lifting, guaranteed preferably consistent reappearance.
B) utilize the typical macromolecular material immobilized antigen such as glutaraldehyde, shitosan, antibody molecule, cost is low, technology maturation.
C) this sensor sheet reveals superior detection sensitivity (lower bound), accuracy, repeatability and stability, and preparation method is ripe simple, is conducive to develop into the product that has a market actual application value and promotes.The utility model utilizes the principle of sonochemistry and micro-stirring effectiveness binding specificity immune response that ultrasonic energy produces and the technology of micro-flowing injection chemiluminescence analysis, improve reaction efficiency, detection sensitivity, degree of accuracy, repeatability and stability are expanded, simplify analytic process, shorten overall detection required time, reduce reagent consumption, further reduce testing cost, detection efficiency and performance are improved, be conducive to high performance price ratio and realize clinical analysis, food security, the online express-analysis in multiple fields such as environmental monitoring.
Accompanying drawing explanation
Fig. 1 is the structural representation of the utility model device;
Fig. 2 is the A-A cross-section separation structure schematic diagram of the utility model device.
Embodiment
Below in conjunction with accompanying drawing, the utility model is further analyzed.
As shown in Figure 1 and Figure 2, the utility model device comprises acoustic energy transmission and chemiluminescence immunoassay reaction checking device part, control test section to embodiment 1..
Described acoustic energy transmission comprises transducer fixing base 111, acoustic energy damping vibration attenuation sheet 110, ultrasonic transducer 101, ultrasonic coupling colloid 102, the integrated glass substrate 103 of sound focusing lens arra, replaceable chemiluminescence immunoassay bio-sensing substrate 104, silicone rubber O-ring 109, optically focused clear glass 107, diaphragm and optical lens 108, optical signal detecting circuit 106 from bottom to up successively with chemiluminescence immunoassay reaction checking device part.
Set gradually from bottom to up and form flow injection reaction tank/injection device by the integrated glass substrate 103 of sound focusing lens arra, silicone rubber O-ring 109, optically focused clear glass 107;
High-frequency transducer 101 is placed between macromolecule ultrasonic coupling colloid 102 and acoustic energy damping vibration attenuation sheet 110, controls the radiation intensity of high-frequency transducer's 101 ultrasonic energies by PID ultrasonic energy mode control module 202.
The integrated glass substrate 103(50mm × 40 × 10mm of sound focusing lens arra) have the import and export of two passages as Fluid Transport, two passages are located at respectively the both sides of ultrasonic coupling colloid 102; Integrated glass substrate 103 upper surfaces of sound focusing lens arra have groove (20mm × 10mm × 2mm), length and width tolerance are ± 1mm, height tolerance is ± 0.5mm to place successively from bottom to up sonac 201, replaceable chemiluminescence immunoassay bio-sensing substrate 104.This groove and two passages form inverted U-shaped structure, make fluid flow to another passage from a passage.
Be provided with silicone rubber O-ring 109 at the integrated glass substrate of sound focusing lens arra 103 edges, for seal flow injection reaction pond/injection device, regulate the height of the flowing reactive chamber 105 of flow injection reaction tank/injection device, and form enclosed construction with optically focused clear glass 107.
The height of the flowing reactive chamber 105 of described flow injection reaction tank/injection device is 1～3mm.
Described replaceable chemiluminescence immunoassay bio-sensing substrate 104 is to form the crosslinked basement membrane of carrier surface by the crosslinked glutaraldehyde of silylating reagent, shitosan etc., form basement membrane, further immobilized antigen molecule (or antibody molecule) and catalyzing enzyme on basement membrane, wherein catalyzing enzyme reacts in order to catalytic oxidation-reduction, produce electroactive material and cause curent change, the kind of conventional enzyme is alkaline phosphatase, horseradish peroxidase etc.
Described optically focused clear glass 107 is greater than unorganic glass or the macromolecule glass of 90 ﹪ for transmittance; Be carved with lensing groove at the lower surface of optically focused clear glass 107, be used for assembling chemiluminescent intensity.By adjusting the degree of depth and the radius of circle size of lensing groove, control sonac 201 sends light signal and makes focus on diaphragm and optical lens 108 surfaces through lensing groove.
In the flowing reactive chamber 105 of flow injection reaction tank/injection device, be provided with replaceable chemiluminescence immunoassay bio-sensing substrate 104, the lower surface of replaceable chemiluminescence immunoassay bio-sensing substrate 104 is stained with sonac 201, and the material of its sonac 201 is Kynoar PVDF.
Replaceable chemiluminescence immunoassay bio-sensing substrate 104 sees through successively optically focused clear glass 107 by fluorescence signal and passes to optical signal detecting circuit 106 with diaphragm and optical lens 108, and photovoltaic reaction occurs, the picking up of settling signal.
Described control test section comprises sonac 201, PID ultrasonic energy mode control module 202, faint optical signal processing module 203, data analysis and feedback control module 204, energy-controlled mode and chemiluminescence immunoassay reaction system database 205.
PID ultrasonic energy mode control module 202 is as controlled processing unit, receive the ultrasonic energy signal of sonac 201 and the feedback signal of energy-controlled mode and chemiluminescence immunoassay reaction system database 205, PID ultrasonic energy mode control module 202 drives ultrasonic transducer 101 to start mode of operation; Faint optical signal processing module 203 receives the collection signal of optically focused clear glass 107, energy-controlled mode and chemiluminescence immunoassay reaction system database 205 are by the signal of data analysis and feedback control module 204 analyzing and processing faint optical signal processing modules 203, then the signal after identification is transmitted to PID ultrasonic energy mode control module 202, finally regulate the ultrasound emission pattern (frequency and intensity) of ultrasonic transducer 101.
Embodiment 2. utilizes above-mentioned device to carry out the analytical approach of chemiluminescence immunoassay biological respinse mensuration.
Step (1). regulate the field of radiational energy of ultrasonic transducer 101
Concave spherical surface size by sound lens on ultrasonic coupling colloid 102 is focused, and making focus is 1～2mm to the vertical range on replaceable chemiluminescence immunoassay bio-sensing substrate 104 surfaces.
On described ultrasonic coupling colloid 102, the characteristic dimension of the concave spherical surface of sound lens is determined by design parameters such as the characteristic dimensions of the flowing reactive chamber 105 of energy size, macromolecule gold film electrode substrate 107 materials and thickness and the flow injection reaction tank/injection device of ultrasonic frequency, focusing.
Overall principle is: 1. regulate ultrasonic transducer 101 intensity to make the interior solution to be measured in flowing reactive chamber 105 of flow injection reaction tank/injection device can produce the disturbance of machinery and temperature, promote diffusional effect, the energy barrier that overcomes reaction, has improved reaction efficiency; 2. be adjusted to suitable intensity, can promote that in solution to be measured, non-specific binding molecular separation between antibody/antigen is reset, and change to specific binding, make solution reaction to be measured abundant.
Step (2). carry out micro-flowing injection analysis
The flowing reactive chamber 105 that solution to be measured, current-carrying are injected into flow injection reaction tank/injection device through a passage constant speed reaches the mixing of the two, finally flows out through another passage.
Carrying out in micro-flowing injection analytic process, if be fixed with antigen molecule on replaceable chemiluminescence immunoassay bio-sensing substrate 104, make the corresponding antibodies generation specific binding reaction in solution to be measured.The microminiaturization of reaction tank has been realized in the flowing reactive chamber 105 of described flow injection reaction tank/injection device by photoetching or etching method.
Step (3). ultrasound emission energy intelligent monitoring process
Carrying out in the process of step (2), the utility model carries out ultrasound emission energy intelligent monitoring simultaneously.Described Based Intelligent Control process comprises two feedback signal path, is respectively feedback signal path, the detection feedback network of ultrasonic energy.
The feedback signal of described ultrasonic energy is detected the intensity of ultrasonic radiation energy by sonac 201, itself and ultrasonic transducer 101 and PID ultrasonic energy mode control module 202 form a closed loop measurement and control system.
PID ultrasonic energy mode control module 202 receives the ultrasonic energy signal of sonac 201, and PID ultrasonic energy mode control module 202 drives ultrasonic transducer 101 to start mode of operation.
Described detection feedback network is to process detection module 203, data analysis and feedback control module 204, energy-controlled mode and chemiluminescence immunoassay reaction system database 205 by PID ultrasonic energy mode control module 202, ultra-weak electronic signal to form closed loop measurement and control system.
Ultra-weak electronic signal is processed detection module 203 and is processed the electrochemical signals that optical signal detecting circuit 106 obtains, regulate the emittance of ultrasonic transducer 101 to determine preferably immune response effect by ultrasonic energy mode control module 202, data analysis and feedback control module 204 will be integrated the parameter sets of different immune response systems, forming energy control model and chemiluminescence immunoassay reaction system database 205, the content of these databases will provide necessary pid control parameter for the application of different immune detection analysis systems.Further, utilize the measured ultrasonic energy feedback parameter of standard Quality Control immune response solution and sonac 201, the dynamic effect of the control parameter that obtains ultrasonic radiation energy to electro-chemistry immunity reaction, by dynamics data between reaction effect parameter and ultrasonic energy control parameter is carried out to matching, obtain the Optimal Parameters for controlling, these Optimal Parameters and pid control parameter carry out information fusion, obtain the concentration of target antibody in solution to be measured or antigen.
Described intelligent monitoring method, according to the feature kinetic curve of the standard immunoassay chemical reaction intensity obtaining in advance, obtains the control parameter of ultrasonic radiation energy, controls chemo-immunity reaction.
The preparation of the replaceable chemiluminescence immunoassay bio-sensing of embodiment 3. substrate 104
The preparation of immunosensor can be divided into two kinds of modes: online preparation and off-line preparation.The utility model is adopted off-line preparation, and online preparation method is the content of another one patent.
(1) determined antigen is dissolved in to buffer solution, selected buffer solution is different because of antigen type, and standard is to make immunoreactive activity and chemiluminescence signal response reach maximum, how to judge and can judge by the comparison of standard channel.
(2) to slide carrier surface carry out pre-service obtain smooth, clean, hydrophilic surface.
(3) configuration finite concentration epoxypropane base trimethyl silane solution, places and it was fully hydrolyzed in 60 minutes, then pipettes 50 microlitre solution and drips in carrier surface, heats 60 minutes at 93 ℃.Drip on the slide processed in epoxypropane base trimethyl silane of 40 microlitre 1% chitosan-acetic acid solutions, then be placed in baking oven to heat 50 minutes under the boiling water condition of 100 ℃, obtain crosslinked with silicane glutaraldehyde (shitosan) basement membrane.
(4) 40 microlitre antigenic solutions are dripped on crosslinked with silicane glutaraldehyde (shitosan) basement membrane, in 4 ℃ of refrigerators, slowly volatilize 8 hours.
(5) drip on (4) step gained basement membrane with 40 microlitre bovine serum albumin solutions, sealing active site, obtains immunologic function film.
(6) above-mentioned immunologic function film is placed in the embedded groove of chemiluminescence flow cell and forms immune bioanalysis pick-up unit.
(7) affect institute's adaptive immune biological function basement membrane and immunosensor performance principal element exist three aspects: (a) time enough and suitable temperature make the abundant hydrolysis reaction of epoxypropane base trimethyl silane; (b) preparation of immunity biosensor is subject to the impact of crosslinked glutaraldehyde (shitosan) surface topography, this depends primarily on the consumption of epoxypropane base trimethyl silane and glutaraldehyde (shitosan) in preparation process, only have consumption proportion suitable, could obtain rule, evenly, high conformity, present the crosslinked basement membrane of mesh-like structure, thereby it is high to prepare stability, the immunologic function film that performance is good; (c) the pH value of buffer solution: only, under certain acidity, antigen just has optimum activity.If acidity departs from this numerical value, the performance of immunologic function basement membrane and sensor will be affected.
The mensuration of embodiment 4. determined antigens: the optimal conditions of (1) immunoassays is the effective actives that guarantee antibody, antigen and marker enzyme, and the reactive conditions of chemiluminescence reality.(2), under optimum determining condition, by the standard solution of variable concentrations antigen or sample box and quantitative enzyme mark antibody solution, after incubation, ultrasound intensity is determined and is controlled curve by Quality Control.Then be injected in the flowing reactive chamber 105 of flow injection reaction tank/injection device by (micro-) flowing injection device, in ultrasonic incubation immune response in early stage, not combined resolvase labeling antibody is caught by fixing antigen/antibody in sensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104), and immune conjugate is taken out of flowing reactive chamber 105 substantially.Inject chemical luminous substrate according to (micro-) flow injection mode of standard and note sensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104), captive enzyme labelled antibody carries out catalytic reaction to chemical luminous system and obtains luminous signal, analyze identification according to the typical curve of measuring in Quality Control process, obtain antigen concentration in sample by the mode of multiparameter information fusion.
The optimization immunoreaction measurement condition of embodiment 5. Quality Control test experiments should comprise following three aspects:
The amount of enzyme labelled antibody in reaction solution: if adopt non-competing immune analysis method, by the online incubation of reaction solution and the ultrasonic agitation transferring energy of determined antigen and fixed amount enzymic-labelled antibody, after immune response finishes, separate enzyme mark sky combination and free and carry by being fixed on antigen on immunosensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104), by the enzyme labelled antibody cataluminescence reaction that is fixed on the antigen capture on immunosensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104), thereby produce signal and reduce the amount of carrying out indirect determination determined antigen.In reaction solution the optimization of the amount of enzyme labelled antibody obtain maximum detection range and the sensitiveest be standard.If the amount of enzyme labelled antibody is less than this value, can make sensing range dwindle; If the amount of enzyme labelled antibody is greater than this value, can make background signal increase, measurement result is less than normal.
After incubation, immune complex flows through the flowing velocity ultrasound intensity of immunosensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104): the time of being detained at sensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104) by reason immune complex is longer, be that flow velocity is slower, the amount of resolvase labeling antibody that is fixed on the antigen capture on immunosensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104) is more; Otherwise the quantity of the catch of resolvase labeling antibody is just few; The former separating effect is relatively better under certain conditions, and the effect that the latter separates is just weaker.But under these two kinds of situations, all can exist and must non-specificly adsorb.Non-specific adsorption may cause noise to improve, and has influence on sensitivity and the accuracy of sensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104).Consider that in addition ultrasonic one side can accelerate the reaction process of antibody/antigen and can reduce on the other hand the background noise that non-specific adsorption causes, the improvement that ultrasonic existence can be larger detects performance, improves detection efficiency.But the improper configuration of ultrasound intensity also may cause the appearance of negative factor, Here it is after intensity is excessive, can cause antibody/antigen inactivation and sensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104) thus the sensitive membrane on surface occurs that the problem coming off has a strong impact on performance.In design of the present utility model owing to having adopted focus ultrasonic lens, can greatly reduce the impact of ultrasonic energy intensity on sensitive membrane, guarantee energy configuration enough in focus ultrasonic district simultaneously, be easy to improve ultrasonic immunoreactive speed and adequacy, further improve and improve immune detection performance.The reaction level of chemistry.Take into account the demand of fast detecting clinically, the flow velocity that after ultrasonic incubation, immune complex flows through immunosensor is optimized with the impact of free enzyme labelled antibody separating effect combination.
Embodiment 6. application examples: testing process is first by testing sample and excessive a little HRP labelled antibody off-line incubation, then this immune potpourri is passed into the flowing reactive chamber 105 of flow injection reaction tank/injection device, in back incubation, unconjugated enzyme labelled antibody is by fixing antigen capture in immunosensor (replaceable chemiluminescence immunoassay bio-sensing substrate 104), and determined antigen-enzyme labelled antibody immune complex is rinsed out.Enzyme labelled antibody based on catching carries out chemiluminescence detection to the catalytic action of luminol-H2O2 chemiluminescence reaction.Typical enzyme labelled antibody is horseradish peroxidase-labeled.
Claims (1)
1. a chemiluminescence immunoassay biology sensor pick-up unit, is characterized in that comprising acoustic energy transmission and chemiluminescence immunoassay reaction checking device part, control test section;
Described acoustic energy transmission comprises transducer fixing base (111) from bottom to up successively with chemiluminescence immunoassay reaction checking device part, acoustic energy damping vibration attenuation sheet (110), ultrasonic transducer (101), ultrasonic coupling colloid (102), the integrated glass substrate of sound focusing lens arra (103), replaceable chemiluminescence immunoassay bio-sensing substrate (104), silicone rubber O-ring (109), optically focused clear glass (107), diaphragm and optical lens (108), optical signal detecting circuit (106),
Set gradually from bottom to up and form flow injection reaction tank/injection device by the integrated glass substrate of sound focusing lens arra (103), silicone rubber O-ring (109), optically focused clear glass (107);
High-frequency transducer (101) is placed between macromolecule ultrasonic coupling colloid (102) and acoustic energy damping vibration attenuation sheet (110), controls the radiation intensity of high-frequency transducer (101) ultrasonic energy by PID ultrasonic energy mode control module (202);
Ultrasonic coupling colloid (102) is made up of ultrasonic coupling agent and macromolecule membrane; The wherein acoustic impedance characteristic of macromolecule membrane and the acoustic impedance characteristic close of selected ultrasonic coupling agent;
Ultrasonic coupling colloid (102) upper surface is provided with acoustic lens array, and for focused ultrasound energy, ultrasonic coupling colloid (102) is close to by acoustic lens array and the integrated glass substrate of sound focusing lens arra (103); The concave spherical surface size of sound lens is determined by focal length, and focal length is the distance of lens surface to the focus of supersonic beam focusing; By adjusting the concave spherical surface size of sound lens, controlling focus is 1～2mm to the vertical range on replaceable chemiluminescence immunoassay bio-sensing substrate (104) surface;
The integrated glass substrate of sound focusing lens arra (103) has the import and export of two passages as Fluid Transport, and two passages are located at respectively the both sides of ultrasonic coupling colloid (102); The integrated glass substrate of sound focusing lens arra (103) upper surface has groove, places successively from bottom to up sonac (201), replaceable chemiluminescence immunoassay bio-sensing substrate (104); This groove and two passages form inverted U-shaped structure, make fluid flow to another passage from a passage;
Be provided with silicone rubber O-ring (109) at the integrated glass substrate of sound focusing lens arra (103) edge, for seal flow injection reaction pond/injection device, regulate the height of the flowing reactive chamber (105) of flow injection reaction tank/injection device, and form enclosed construction with optically focused clear glass (107);
The height of the flowing reactive chamber (105) of described flow injection reaction tank/injection device is 1～3mm;
Described replaceable chemiluminescence immunoassay bio-sensing substrate (104) is to form the crosslinked basement membrane of carrier surface by the crosslinked glutaraldehyde of silylating reagent, shitosan, form basement membrane, further immobilized antigen molecule or antibody molecule and catalyzing enzyme on basement membrane, wherein catalyzing enzyme reacts in order to catalytic oxidation-reduction, produces electroactive material and cause curent change;
Described optically focused clear glass (107) is greater than unorganic glass or the macromolecule glass of 90 ﹪ for transmittance; Lower surface at optically focused clear glass (107) is carved with lensing groove, is used for assembling chemiluminescent intensity; By adjusting the degree of depth and the radius of circle size of lensing groove, control sonac (201) sends light signal and makes focus on diaphragm and optical lens (108) surface through lensing groove;
In the flowing reactive chamber (105) of flow injection reaction tank/injection device, be provided with replaceable chemiluminescence immunoassay bio-sensing substrate (104), the lower surface of replaceable chemiluminescence immunoassay bio-sensing substrate (104) is stained with sonac (201), and the material of its sonac (201) is Kynoar PVDF;
Replaceable chemiluminescence immunoassay bio-sensing substrate (104) sees through successively optically focused clear glass (107) and diaphragm and optical lens (108) by fluorescence signal and passes to optical signal detecting circuit (106), and photovoltaic reaction occurs, the picking up of settling signal;
Described control test section comprises sonac (201), PID ultrasonic energy mode control module (202), faint optical signal processing module (203), data analysis and feedback control module (204), energy-controlled mode and chemiluminescence immunoassay reaction system database (205);
PID ultrasonic energy mode control module (202) is as controlled processing unit, receive the ultrasonic energy signal of sonac (201) and the feedback signal of energy-controlled mode and chemiluminescence immunoassay reaction system database (205), PID ultrasonic energy mode control module (202) drives ultrasonic transducer (101) to start mode of operation; Faint optical signal processing module (203) receives the collection signal of optically focused clear glass (107), energy-controlled mode and chemiluminescence immunoassay reaction system database (205) are by the signal of data analysis and feedback control module (204) analyzing and processing faint optical signal processing module (203), then the signal after identification is transmitted to PID ultrasonic energy mode control module (202), finally regulate the ultrasound emission pattern of ultrasonic transducer (101).
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103604921A (en) * | 2013-11-05 | 2014-02-26 | 浙江大学 | Chemiluminiscence immuno biosensor detection device and detection analysis method |
| CN103743723A (en) * | 2014-01-14 | 2014-04-23 | 中国人民解放军63750部队后勤部防检环监所 | High sensitivity bioluminescence detector |
| CN106153951A (en) * | 2016-06-17 | 2016-11-23 | 天津智巧数据科技有限公司 | The visualization monomolecular detection method of Gibberella zeae alcohols in a kind of milk product |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103604921A (en) * | 2013-11-05 | 2014-02-26 | 浙江大学 | Chemiluminiscence immuno biosensor detection device and detection analysis method |
| CN103743723A (en) * | 2014-01-14 | 2014-04-23 | 中国人民解放军63750部队后勤部防检环监所 | High sensitivity bioluminescence detector |
| CN103743723B (en) * | 2014-01-14 | 2016-01-13 | 中国人民解放军63750部队后勤部防检环监所 | A kind of high sensitivity bioluminescence detector |
| CN106153951A (en) * | 2016-06-17 | 2016-11-23 | 天津智巧数据科技有限公司 | The visualization monomolecular detection method of Gibberella zeae alcohols in a kind of milk product |
| CN108760688A (en) * | 2018-02-13 | 2018-11-06 | 中国人民解放军91054部队 | A kind of detection and identification device of underwater micro- trace chemical |
| CN108760688B (en) * | 2018-02-13 | 2021-12-10 | 中国人民解放军91054部队 | Detection and identification device for underwater micro-trace chemical substances |
| CN109507008A (en) * | 2018-10-24 | 2019-03-22 | 西安交通大学 | A kind of microlayer model snap cure device based on surface acoustic wave drop micro-fluidic chip |
| CN109507008B (en) * | 2018-10-24 | 2020-02-14 | 西安交通大学 | Micro-droplet rapid solidification device based on surface acoustic wave droplet micro-fluidic chip |
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