CN208066375U - A kind of electro-chemistry immunity micro-fluidic detection card - Google Patents
A kind of electro-chemistry immunity micro-fluidic detection card Download PDFInfo
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- CN208066375U CN208066375U CN201721858225.6U CN201721858225U CN208066375U CN 208066375 U CN208066375 U CN 208066375U CN 201721858225 U CN201721858225 U CN 201721858225U CN 208066375 U CN208066375 U CN 208066375U
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Abstract
The utility model provides a kind of electro-chemistry immunity micro-fluidic detection card, the detection card includes hydrophilic upper cover, middle interlayer and insulating base, contain at least a pair of of test electrode and at least one detection reagent spraying area on the insulating base, the test electrode is connected with contact conductor, the contact conductor is connected with the detection module of test equipment, the test electrode print is on insulating base, the hydrophilic upper cover is covered on middle interlayer, the middle interlayer is covered on insulating base, and the middle interlayer is equipped with microfluidic channel.The utility model can not only allow to need to can be completed on a chip in the work that a laboratory is completed originally, and enormously simplify the manufacturing process of micro-fluidic chip and greatly reduce the manufacturing cost of micro-fluidic chip, in the sensitivity of detection, accuracy, test be linear and test CV etc. obtains ideal effect.
Description
Technical field
The utility model is related to field of biosensors more particularly to rapid in-vitro diagnostic fields, and in particular to Yi Zhong electricity
Chemo-immunity micro-fluidic detection card.
Background technology
In biosystem, there is various substances, they have very the various aspects of biological process
Important influence, it is always the target that analysis science is pursued to carry out fast and accurately analysis to these substances.Environmental monitoring, poison
Also very there is an urgent need to establish various analysis methods quickly, easy for the application fields such as product monitoring and clinical diagnosis.Traditional
Analysis method is based on chemical method, it is often necessary to a series of cumbersome operating process, analytical cycle is long, and sensitivity it is low,
Poor selectivity, poor accuracy.With the development of multi-crossed disciplines area research, new analysis biology techniques-electrochemica biological
Sensor and microfluidic chip analysis technology are born.
Electrochemica biological sensor is the biosensor to come out earliest.In the 1960s, Clark and Lyons are for the first time
It proposes to help a small child urinate by holding his legs apart using the film containing enzyme and is converted into product with glucose, the imagination detected using pH electrodes or oxygen electrode；?
1967, Updike and Hicks were fixed to the polyacrylamide gel film containing glucose oxidase on oxygen electrode and are prepared for
First glucose sensor, has started the history of electrochemica biological sensor.It is sensitive primitive with active material, with electrochemistry
Electrode is signal converter, and the biosensor that signal is detected characterized by current potential, electric current or capacitance is known as electrochemica biological biography
Sensor.
Micro-fluidic chip (Microfluidic chip), also known as Microfluid based Lab on a chip or chip lab (Lab-
On-a-chip), it is to be proposed by the Widmer and Manz of Switzerland in the 1990s, is related in the fields such as biochemistry
Sample preparation, reaction, separation, detection and cell culture, sorting, cracking etc., and these basic operation units are passed through into microcomputer
Power technology and mechanical & electrical technology (MEMS/NEMS) of receiving integrate or are integrated into substantially the chip of one piece several square centimeters (or even smaller)
Up, network is formed using microchannel, controlled fluid runs through whole system, to substitute various work(in conventional biochemical laboratory
Energy instrument and a kind of technology platform of operation.
But traditional micro-fluidic chip processing technology is there are cost height, chip structure is complicated, external driving is needed to manipulate
The defects of fluid flows.And this programme design micro-fluidic chip have it is at low cost, simple in structure, without it is external driving rely on rainbow
Dynamic flow is inhaled, many drawbacks of traditional microfluidic chip is solved, is with a wide range of applications.
Invention content
The purpose of the present invention is overcoming the deficiencies of the prior art and provide a kind of electro-chemistry immunity micro-fluidic detection card, solve
Technical problem of the existing technology：Traditional biochemical immunity analysis usually needs a series of cumbersome operating process, analysis week
Phase is long, and sensitivity is low, selectivity and poor accuracy.Micro-fluidic chip immunological technique to need in a laboratory originally
The work of completion can be completed on a chip now, and the consumption of Biosample and reagent is not only made to be significantly reduced to microlitre very
To nanoliter level, and make the raising of hundred times of analyze speed tenfold, it can to become for the family oriented of realization assay laboratory
Can, but traditional micro-fluidic immuno-chip uses micro electro mechanical processing technology, relative to technologies such as traditional immunochromatography, ELISA
There are still the higher defects of cost；In addition, that there is also structures and manufacturing process is also more complex, it is external to need for traditional microfluidic chip
Driving is come the defects of manipulating fluid flowing.
In order to solve higher traditional microfluidic chip cost, complex manufacturing process, external driving be needed to manipulate fluid stream
The defects of dynamic, We conducted further investigations.As a result, it has been found that by being die cut on the middle interlayer with two-sided mucigel or
The microfluidic channel that makes of modes of other cuttings, and with test electrode insulating base and with the upper of hydrophilic nmature
Immune micro-fluidic chip made of lid assembling can be very good to solve the above problems, and in detection sensitivity, accuracy, p-wire
Property and test CV etc. obtain ideal effect, so as to complete the utility model.
The technical solution of the utility model is as follows：
The electro-chemistry immunity micro-fluidic detection card of the utility model, including hydrophilic upper cover, middle interlayer and insulating base, exhausted
It is connected with electrode containing at least a pair of of test electrode and at least one detection reagent spraying area, the test electrode on edge bottom plate and draws
Line, contact conductor are connected with the detection module of test equipment, and test electrode need to be by way of printing or other modes are made in advance
Make on insulating base.Insulating base with test electrode is covered with middle interlayer, containing by the side of being pre-designed on interlayer in this
The microfluidic channel that case is made；Middle interlayer overlying is stamped one layer of hydrophilic upper cover.The hydrophilic upper cover is equipped with well and row
Stomata.
Material used in the test electrode and contact conductor is conductive material, attached by silk-screen printing or other modes
It on insulating base, the mode of preferably silk-screen printing makes.
The detection reagent spraying area sprays the material formula composition of specific objective ingredient in promising detection blood, should
Compound composition can spray up before middle interlayer is pasted on insulating base to be pasted on insulating base in middle interlayer
It sprays up later, the scheme preferably sprayed up after middle interlayer is pasted on insulating base.
The middle interlayer contain viscosity lyophobic dust, ingredient can be silica gel, polyacrylate or other at
Divide, preferably silica gel or polyacrylate.
Containing according to the microfluidic channel for being pre-designed scheme making on the middle interlayer, microfluidic channel can use
The modes such as cross cutting, laser cutting make, wherein it is preferred that the mode being die cut.
The hydrophilic upper cover of detection card can usually be made by hard or soft material.It is examined for convenient and cost is cut
Consider, the preferably material of flexible plastic material, such as PET, PVC, PC, wherein it is preferred that the soft plastic material of PET materials.
The hydrophily of the hydrophilic upper cover can uniformly spray hydrophilic substance to be made on upper lid,
It can be made by means such as plasma treatment or sided corona treatments；It, preferably will be close in order to ensure the hydrophilic stabilization of upper cover
Aqueous substance even application is in the scheme of upper cover.
The micro-fluidic detection card needs made are cooperateed with necessary instrument to be detected, and instrument can pass through measurement
The variation of the electric signal (such as electric current, resistance, conductance and electrical impedance) after electrode zone reacts is tested to detecting sample
In target substance carry out measured either qualitatively or quantitatively.
Being made by for the electro-chemistry immunity micro-fluidic detection card of the utility model first will be with two-sided mucigel
Middle interlayer is cut into the microfluidic channel of needs by pre-designed scheme, and the middle interlayer of well cutting is then pasted on band
It on the insulating base for having test electrode, then is pasted on middle interlayer and carries hydrophilic upper cover, finally cut into required big
Small and shape, as complete detection card.Specific manufacturing process is as follows：
(1) select a kind of any surface finish, it is non-conductive, to plastic material of the sample without permeability and absorbability as test strips
Insulating base, the material of the plastics can be PET, PVC, PE, PP, PS, ABS etc., preferably PET.
(2) by testing electrode and electrode cable and drying in silk-screen printing or other modes attachment on insulating base,
To ensure electrode and conducting wire adhesion-tight.It can be detected with cross cross-hatching to ensure that the firmness of attachment meets the requirements.It makes
The material of electrode and electrode cable is conductive material, such as carbon and gold, silver, platinum precious metal material, preferably gold.
(3) formulatory agents of test are formulated for, formulatory agents, which contain, to be occurred to be immunized with target substance in detection sample
The antibody of reaction, antibody can be that how anti-monoclonal antibody can also be；Antibody can be marked with label probe and can not also be marked；Label
It can also be conductive that probe, which can be non-conductive,.The preferred conductive colloid gold label probe labelled antibody of this programme, so as to
Amplification test signal is played, the sensitivity of detection is improved.
(4) select it is a kind of containing viscosity lyophobic dust middle interlayer, ingredient can be silica gel, polyacrylate or its
His ingredient, preferably silica gel and polyacrylate.Interlayer is made by insulating material in this, and material can be PET, PC, PVC
Deng wherein it is preferred that the material of soft PET materials.Middle spacer width should be less than or the width equal to insulating base, length should be less than
The length of insulating base is to ensure that electrode cable can be connected to ELISA test strip instrument.
(5) selected middle interlayer is cut with designed scheme.
(6) well cutting is pasted on the middle interlayer of microfluidic channel on insulating base, the material of insulating base can be with
It is PET, PC, PVC etc., wherein it is preferred that the material of PET materials.
(7) bottom plate for pasting middle interlayer is flattened with roll squeezer, during the bubble generated in paste process is squeezed out and made
Interlayer is tight with base plate sticking, avoids the leakage of sample occur when carrying out sample-adding reaction.
(8) the reagent spraying area of the insulating base of interlayer sprays the formulatory agents for test with instrument in pasting
And it is dry, the reagent layer shape being attached to after dry on test electrode is preferably regular shape.
(9) hydrophilic upper cover is pasted on the insulating base for sprayed detection reagent.The hydrophily of hydrophilic upper cover can be
Hydrophilic substance is uniformly sprayed in what is made on upper lid, can also be by means such as plasma treatment or sided corona treatments
It makes；In order to ensure the hydrophilic stabilization of upper cover, preferably by hydroaropic substance even application in the scheme of upper cover；Hydrophilic upper cover
It can usually be made by hard or soft material.For observation and cut convenient and cost consideration, preferably soft transparent plastics material
The material of matter, such as PET, PVC, PC, wherein it is preferred that the soft transparent plastic material of PET materials.
(10) insulating base for pasting hydrophilic upper cover is flattened with roll squeezer, the bubble generated in paste process is squeezed out
And it is tight so that hydrophilic upper cover is pasted with middle interlayer, avoids the leakage of sample occur when carrying out sample-adding reaction.
(11) the big plate of detection card for pasting hydrophilic upper cover cuts into required size and shape with cutting machine, as at product examine
Survey card.
Evaluation scheme：
(1) blood sample is acquired, blood sample is adjusted to the sample of at least five test concentrations with object to be measured substance.
(2) the blood sample sample of each concentration to be measured is divided into bisection.First part is detected with standard detection instrument in blood sample
The content of target substance；Second part of immune micro-fluidic detection made of this programme is stuck on mating detecting instrument and is detected.
(3) compare the immune micro-fluidic detection card blood sample test result made of this programme and reference instrument testing result
Deviation, and calculate test CV and test linearly.
The utility model has the following technical effect that：The utility model in view of the problems existing in the prior art, provides one
The design scheme of immune micro-fluidic detection card of the kind based on biosensor technology.With this solution, can not only make to need originally
The work to be completed in a laboratory can be completed on a chip, and enormously simplify the system of micro-fluidic chip
Make process and greatly reduce the manufacturing cost of micro-fluidic chip, in the sensitivity of detection, accuracy, test is linear and tests
CV etc. obtains ideal effect.According to the utility model, it is quickly cheap that we realize micro-fluidic chip high-volume
Make, the immune micro-fluidic chip of making for testing CRP blood samples, CRP test the range of linearity up to 0.5mg/L~
200mg/L, test CV are less than 10%, are controlled within 10% range with the deviations in accuracy for compareing instrument.
Description of the drawings
Fig. 1 is the utility model electro-chemistry immunity micro-fluidic detection card overall appearance figure.
Fig. 2 is the insulating base figure that electro-chemistry immunity micro-fluidic detection card printing has test electrode.
Fig. 3 is the middle interlayer schematic diagram with microfluidic channel.
Fig. 4 is hydrophilic upper cover schematic diagram.
1-1 is detection card well in figure, and 1-2 is detection reagent spraying area, and 1-3 is test reaction detection zone, and 2-1 is exhausted
Edge bottom plate, 2-2 are test electrode, and 2-3 is contact conductor, and 3-1 is middle interlayer, and 3-2 is microfluidic channel, and 4-1 is hydrophilic upper cover,
4-2 is gas vent.
Specific implementation mode
The utility model is explained in detail with reference to specific embodiment, but the technical scope of the utility model is not limited to
These embodiments.
The electro-chemistry immunity micro-fluidic detection card of the utility model, including hydrophilic upper cover, middle interlayer and insulating base, described
Insulating base be equipped with detection reagent spraying area, test reaction detection zone, test electrode and contact conductor, the electrode draw
Line is connected with the detection module of test equipment, and the test electrode print is on insulating base, hydrophilic upper cover covering
On middle interlayer, the middle interlayer is covered on insulating base, and the middle interlayer is equipped with microfluidic channel；The parent
It is waterborne to be covered with well and gas vent.
The preparation process of the electro-chemistry immunity micro-fluidic detection card of the utility model is as follows：
(1) select a kind of any surface finish, it is non-conductive, to PET insulating base of the sample without permeability and absorbability.
(2) upper test electrode and electrode cable are printed by way of silk-screen printing on insulating base and is dried.It makes
The material of electrode and electrode cable is gold.
(3) formulatory agents of test are formulated for.Reagent set becomes：10% (W/V) sodium carboxymethylcellulose, 3% (W/V)
Hydroxyethyl cellulose, 0.5% (W/V) BSA, 0.1M pH 7.0PBS buffer solutions, the CRP that 1mg/mL is marked with colloidal gold probe are more
It is anti-.
(4) a kind of middle interlayer of the hydrophobic polyacrylate containing viscosity is selected, interlayer is made by insulating material in this,
Its material is the material of soft PET materials.Middle spacer width is equal to the width of insulating base, length is less than the length of insulating base
To ensure that electrode cable can be connected to ELISA test strip instrument.
(5) selected middle interlayer is die cut with designed mold.
(6) middle interlayer of the well cutting with microfluidic channel is pasted on the insulating base of PET materials.
(7) bottom plate for pasting middle interlayer is flattened with roll squeezer, during the bubble generated in paste process is squeezed out and made
Interlayer is tight with base plate sticking, avoids the leakage of sample occur when carrying out sample-adding reaction.
(8) the reagent spraying area of the insulating base of interlayer sprays the formulatory agents for test with instrument in pasting
And it is dry, the reagent layer appearance being attached to after dry on test electrode should uniform rule.
(9) hydrophilic upper cover is pasted on the insulating base for sprayed detection reagent.The hydrophily of hydrophilic upper cover is will be close
Aqueous substance is uniformly sprayed in what is made on upper lid, and hydrophilic upper cover is the soft plastic material of transparent PET materials.
(10) insulating base for pasting hydrophilic upper cover is flattened with roll squeezer, the bubble generated in paste process is squeezed out
And it is tight so that hydrophilic upper cover is pasted with middle interlayer, avoids the leakage of sample occur when carrying out sample-adding reaction.
(11) the big plate of detection card for pasting hydrophilic upper cover cuts into required size and shape with cutting machine, as at product examine
Survey card.
Assess the immune micro-fluidic detection card detection CRP blood samples detection sensitivity of the utility model design, accuracy,
The effect for testing linear and test CV etc., is as follows：
(1) acquire blood sample, with CRP antigen standards adjust blood sample in CRP concentration, by blood sample be adjusted to 0.5mg/L,
The sample of six concentration of 10mg/L, 50mg/L, 100mg/L, 150mg/L and 200mg/L.
(2) the blood sample sample of each concentration is divided into bisection.First part is detected with control test instrument；Second part with originally
The micro-fluidic detection that scheme makes, which is stuck on mating detecting instrument, to be detected, this exemplary instrument is examined using AC impedence method
It surveys.
(3) compare the deviation and linear correlation of the detection card test result and reference instrument testing result that are made of this programme
Property.
(4) CV for the detection card testing result that calculating this programme makes.
Its testing result is as follows：
The micro-fluidic detection card test result that the control instrument of table 1 and this programme make
It can be seen that from the test result of table 1：When the immune micro-fluidic detection that this programme makes is stuck in detection CRP blood samples
Compared with control instrument, there is good linear dependence between the two；The detection card detection sensitivity that this programme makes can
Reach 0.5mg/L, test CV is respectively less than 10%, is also respectively less than 10% with the deviations in accuracy for compareing instrument, shows excellent
Test performance.
Claims (10)
1. a kind of electro-chemistry immunity micro-fluidic detection card, which is characterized in that the detection card includes hydrophilic upper cover（4-1）, in
Interlayer（3-1）And insulating base（2-1）, the insulating base（2-1）It is upper to contain at least a pair of of test electrode（2-2）At least
One detection reagent spraying area（1-2）, the test electrode（2-2）It is connected with contact conductor（2-3）, the contact conductor
（2-3）It is connected with the detection module of test equipment, the test electrode（2-2）It is printed on insulating base（2-1）On, it is described
Hydrophilic upper cover（4-1）It is covered in middle interlayer（3-1）On, the middle interlayer（3-1）It is covered in insulating base（2-1）On, it is described
Middle interlayer（3-1）It is equipped with microfluidic channel（3-2）.
2. electro-chemistry immunity micro-fluidic detection card according to claim 1, which is characterized in that the test electrode（2-
2）And contact conductor（2-3）Material be conductive material, be attached on insulating base by silk-screen printing.
3. electro-chemistry immunity micro-fluidic detection card according to claim 1, which is characterized in that the detection reagent spraying
Area（1-2）It is coated with the material formula composition of target component in detection blood, which is pasted on absolutely in middle interlayer
Edge bottom plate（2-1）It sprays up before or in middle interlayer（3-1）It is pasted on insulating base（2-1）It sprays up later.
4. electro-chemistry immunity micro-fluidic detection card according to claim 1, which is characterized in that the middle interlayer（3-1）
Containing sticking lyophobic dust, the lyophobic dust is silica gel or polyacrylate.
5. electro-chemistry immunity micro-fluidic detection card according to claim 1, which is characterized in that micro- on the middle interlayer
Flow control channel is made using being die cut or being cut by laser mode.
6. electro-chemistry immunity micro-fluidic detection card according to claim 1, which is characterized in that the hydrophilic upper cover（4-
1）Material be hard or soft material.
7. electro-chemistry immunity micro-fluidic detection card according to claim 6, which is characterized in that the hydrophilic upper cover（4-
1）Material be flexible plastic.
8. electro-chemistry immunity micro-fluidic detection card according to claim 7, which is characterized in that the flexible plastic is
PET, PVC or PC.
9. electro-chemistry immunity micro-fluidic detection card according to claim 1, which is characterized in that the hydrophilic upper cover（4-
1）It is to uniformly spray hydrophilic substance to make on upper lid, or make by plasma treatment or sided corona treatment.
10. electro-chemistry immunity micro-fluidic detection card according to claim 1, which is characterized in that the micro-fluidic detection
Card needs to cooperate with necessary instrument to be detected, and instrument tests the variation of the electric signal after electrode zone reacts by measurement
Target substance in detection sample is carried out measured either qualitatively or quantitatively.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201721858225.6U CN208066375U (en) | 2017-12-27 | 2017-12-27 | A kind of electro-chemistry immunity micro-fluidic detection card |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721858225.6U CN208066375U (en) | 2017-12-27 | 2017-12-27 | A kind of electro-chemistry immunity micro-fluidic detection card |
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| CN201721858225.6U Active CN208066375U (en) | 2017-12-27 | 2017-12-27 | A kind of electro-chemistry immunity micro-fluidic detection card |
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