CN109030812A - A kind of micro-fluidic chip based on immune detection and biochemistry detection, detector and detection method - Google Patents
A kind of micro-fluidic chip based on immune detection and biochemistry detection, detector and detection method Download PDFInfo
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- CN109030812A CN109030812A CN201810794892.5A CN201810794892A CN109030812A CN 109030812 A CN109030812 A CN 109030812A CN 201810794892 A CN201810794892 A CN 201810794892A CN 109030812 A CN109030812 A CN 109030812A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
Abstract
The present invention provides a kind of micro-fluidic chip based on immune detection and biochemistry detection, detector and detection method, which includes substrate, and sample injection unit, immune detection area and biochemistry detection area is arranged on substrate；Immune detection area includes the first reaction member and devil liquor recovery unit, is fixed with the first solidification oxidant layer in the first reaction member；Biochemistry detection area includes the second reaction member, is fixed with the second solidification oxidant layer in the second reaction member；Sample injection unit is connected to the first reaction member and the second reaction member respectively by microchannel.The detector integrates two distinct types of detection on a single die, and structure is simple, is capable of the flowing of accurate control of fluid, and detection is fast and accurate.
Description
Technical field
The invention belongs to detection device and method and technology fields, are related to a kind of miniflow based on immune detection and biochemistry detection
Control chip, detector and detection method.
Background technique
The judgement of disease needs that different information is combined to combine, such as biochemistry detection, immune detection and Image detection.At present absolutely
Most of detection device is all that orientation does a certain detection, such as Biochemical Analyzer, and immunity analysis instrument is all to be with a certain substance
Detect substance.In addition, existing detection device volume is larger, expensive, the more difficult burden of typically small basic hospital.
Chemiluminescence immune analysis method is due to the advantages of it is highly sensitive, high specific, and substitution radiation substantially is exempted from
Epidemic disease analysis and ELISA become a kind of most immunoassay methods of immunoassay diagnostic field application.
There are mainly two types of chemical illumination immunity analysis instruments currently used in the market, and it is immune that one is Full-automatic chemiluminescences
Analyzer, another kind are semi-automatic chemical illumination immunity analysis instruments.
Full-automatic instrument realizes the automatic processing of entire chemiluminescence immune assay process, including sample introduction, sample-adding plus
Reagent, incubation, cleaning, Magneto separate and the detection etc. that shines, test process are participated in without artificial, and test flux is higher, is suitable for
The big large hospital of sample throughput.But full automatic device structure is complicated, and it is bulky, it takes up a large area, equipment manufacturing cost
Height, detection speed are slow.
Semi-automated instrument then only remains chemiluminescence detection and simple Mechanical course structure, remaining process is by artificial
It completes.Instrument cost is greatly reduced, but increases manual operation error, affects the accuracy of result.Such as
The disclosed magnetic microparticle chemiluminescence micro-fluidic chip and CN105842468A for whole blood sample detection of CN105241871A is public
The micro-fluidic chemiluminescence immunoassay detection device opened.
Biochemistry detection is also one of common detection method, doctor can be helped to understand the physical condition of patient, sometimes
It can also check latent disease, be treated in time.
It is mainly sample flow that immune detection and biochemistry detection, which are integrated in the technical problem faced on lesser chip,
Accurate control.
CN107091936A discloses a kind of chemiluminescence immunoassay disc based on microflow control technique, including adding mouth, examination
Agent mouth, detection slot and lead-over groove；Compared with the detection slot, rotation center of the lead-over groove farther away from the disc；First is logical
The one end in road is connected to the end far from the rotation center of the detection slot, and the other end is connected to the lead-over groove, and centre is curved
Folding part；The minimum radius of turn of the first passage is less than the maximum radius of turn of the detection slot；Compared with the lead-over groove,
Rotation center of the waste liquid tank farther away from the disc；The lead-over groove is connected to the waste liquid tank.But its adding mouth, reagent
It is directly connected between mouth and detection slot, does not utilize the accurate control of fluid.
Summary of the invention
It is a kind of based on immune detection and biochemistry detection in view of the deficiencies of the prior art, the present invention intends to provide
Micro-fluidic chip, detector and detection method, the micro-fluidic chip can be compatible with immune detection and biochemistry detection, by two kinds
Different types of detection is integrated on a single die, can be quickly obtained accurate result by an instrument, structure is simple, can
The flowing of accurate control of fluid.
In the present invention unless otherwise specified described " first ", " second ", " third ", " the 4th ", " the 5th " and " the 6th " etc.
Just for the sake of distinguishing each structure, it is not offered as quantity.
The microchannel refers to the channel having a size of 0.01-5.00mm, as 0.05mm, 0.10mm, 0.20mm, 0.30mm,
0.50mm, 1.00mm, 2.00mm, 2.50mm, 3.50mm or 4.50mm etc..
Immune detection area and biochemistry detection area of the present invention are not a specific structure, are intended merely to distinguish
Region where immune detection and biochemistry detection on substrate, the region of the two can also intersect in actual use.
To achieve this purpose, the present invention adopts the following technical scheme:
One of the objects of the present invention is to provide a kind of micro-fluidic chip based on immune detection and biochemistry detection, including base
Sample injection unit, immune detection area and biochemistry detection area are arranged on substrate for plate；
Sample injection unit includes sample introduction tank；
Immune detection area includes the first reaction member and devil liquor recovery unit, and the first reaction member and devil liquor recovery unit are logical
Cross microchannel connection；First reaction member includes the first reactive tank, is fixed with the first solidification oxidant layer in the first reaction member；Waste liquid
Recovery unit includes waste liquid tank；
Biochemistry detection area includes the second reaction member, and the second reaction member includes the second reactive tank, in the second reaction member
It is fixed with the second solidification oxidant layer；
Sample injection unit is connected to the first reaction member and the second reaction member respectively by microchannel.
Micro-fluidic chip provided by the invention will test reagent and be embedded in the first reaction member and the second reaction member in advance
In, it only needs that sample to be detected is added in detection process, it is easy to operate.
Micro-fluidic chip provided by the invention is based on centrifugation technique.When substrate centrifugation center refers to substrate centrifugation
Center.Micro-fluidic chip can be fixed on centrifugal device by the centrifugation to carry out.
First reaction member is connected to devil liquor recovery unit by being provided with the microchannel of third valve.
Preferably, the sample injection unit is anti-with the first reaction member and second respectively by the microchannel for being provided with the first valve
Unit is answered to be connected to.
The valve of its microchannel of micro-fluidic chip provided by the invention setting is capable of the flowing of accurate control of fluid.
The sample injection unit further includes the first quantitative slot and collecting tank, and sample introduction tank, the first quantitative slot and collecting tank successively lead to
Cross microchannel connection；
Preferably, the first quantitative slot reacts single by being equipped with the microchannel of the first valve with the first reaction member and second respectively
Member connection.First quantitative slot quantifies sample using cavity volume, keeps testing result more accurate.When the sample that sample introduction tank is added is
When whole blood, whole blood sample blood plasma or serum after being centrifuged can be stayed in the first quantitative slot after substrate rotation, and erythrocyte can enter
Collecting tank.
Preferably, the sample injection unit further includes the first distributing trough, and the first quantitative slot is by the first distributing trough respectively with the
One reaction member and the connection of the second reaction member；
Preferably, first distributing trough is set between the first valve and the first reaction member or the second reaction member.
The setting of first distributing trough is respectively arranged with multiple first reaction members convenient for immune detection area and biochemistry detection area
With the control of fluid when the second reaction member.
Preferably, first distributing trough is arc-shaped or rectangular etc., those skilled in the art can according to actual needs into
Row selection.
The immune detection area also sets up the second quantitative slot, and the second quantitative slot pass through respectively microchannel and sample injection unit and
The connection of first reaction member；
Preferably, second quantitative slot is set between the first valve and the first reaction member；
Preferably, the second valve, the second valve and substrate are set on the microchannel that the second quantitative slot is connected to the first reaction member
The distance at centrifugation center less than microchannel where the second valve remainder at a distance from the center of centrifugation.The effect of second quantitative slot
It is the sample volume of quantitative reaction, keeps reaction result more accurate.
Preferably, second quantitative slot is set between the first distributing trough and the first reaction member.
Preferably, the immune detection area also sets up the second allocation unit, the second allocation unit respectively with the second quantitative slot
It is connected to the first reactive tank.When immune detection area carries out panimmunity detection simultaneously, second distributing trough is used for each
Sample to be tested is distributed in first reaction member.
Preferably, second allocation unit is set between the second valve and the first reaction member.
Preferably, second allocation unit includes the second distributing trough.
Preferably, second distributing trough is arc-shaped or rectangular etc., those skilled in the art can according to actual needs into
Row selection.
The immune detection area further includes the first liquid storage unit, the first liquid storage unit and the first reaction member
And/or second allocation unit by microchannel be connected to, be preferably connected to the second allocation unit by microchannel.First liquid
Storage unit is used to provide required liquid to immune detection area.
Preferably, first liquid storage unit includes at least one fluid storage tanks, such as 2,3,5,8,
12,18 or 20 etc..The fluid storage tanks include liquid storage tank, and the inside of the liquid storage tank, which is equipped with, accommodates liquid
The cavity of body reagent；First sealing-plug, the first sealing-plug are detachably connected with the liquid storage tank, and can be by the sky
One end of chamber seals；The other end of cavity can be connected to by microchannel with the first reaction member and/or the second allocation unit；And
Hot melt sealing-plug is set on the microchannel that cavity is connected to the first reaction member and/or the second allocation unit.Need release liquid
When body, the hot melt sealing-plug of fluid storage tanks is melted using laser or the mode of heating.The immune detection area
The liquid of fluid storage tanks storage can be dilution property liquid, substrate solution and cleaning solution etc..
First reaction member further includes third quantitative slot and the first mixing channel, and sample to be tested can be according to by microchannel
It is secondary to pass through third quantitative slot, the first mixing channel and the first reactive tank.
Setting solidifies oxidant layer in first mixing channel or the first reactive tank, and curing agent is arranged in preferably the first mixing channel
Layer.The effect of third quantitative slot is the sample volume of quantitative reaction, keeps reaction result more accurate.Sample to be tested elder generation and curing agent
Layer is mixed, and is entered the first reactive tank later and is reacted.
Preferably, the 4th valve is set on the microchannel that third quantitative slot is connected to the first mixing channel.
Preferably, the third quantitative slot is also connected to liquid storage unit by microchannel.The third quantitative slot can
It is connected to dilution storage tank, for diluting sample to be tested.
Preferably, the immune detection area includes multiple first reaction members, such as 2,3,5,8,10,15
A, 18,25 or 30 etc., the multiple first reaction member is connected to the second allocation unit respectively.First reaction member
Number can be determined according to detection.
Preferably, the first curing agent layer choosing self-curing reagent layer.
Preferably, the sample injection unit, the second quantitative slot, the first reaction member and the centrifugation of devil liquor recovery cell distance substrate
Neutral distance successively increases.
The biochemistry detection area also sets up the 4th quantitative slot, and sample injection unit connects by the 4th quantitative slot and the second reaction member
It is logical.
Preferably, the 4th quantitative slot is set between the first valve and the second reaction member, and the 4th quantitative slot and
The 5th valve is set on the microchannel of two reaction members connection.The effect of 4th quantitative slot is the sample volume of quantitative reaction, is made anti-
Answer result more accurate.
Preferably, the 4th quantitative slot is connected to the first distributing trough.
The biochemistry detection area also sets up the second mixed cell, and sample injection unit reacts single with second by the second mixed cell
Member connection, the second mixed cell are set between the first valve and the second reaction member.Second mixed cell is used for sample to be tested
It is mixed with dilution etc..
Preferably, the 6th valve is set on the microchannel that the second mixed cell is connected to the second reaction member.
Preferably, first valve, the second valve, third valve, the 4th valve, the 5th valve and the 6th valve are independently selected from drain valve
And/or capillary valves.
Drain valve is that microchannel surface is carried out hydrophobically modified, and such microchannel surface tension greatly increases, when liquid exists
When passing through this microchannel under external force, required external force increases therewith.Chip is shown as when applying compared under the slow-speed of revolution, liquid
The centrifugal force that body is subject to is smaller, and liquid can not be by this microchannel, and when revolving speed improves to a certain extent, it is micro- that liquid can break through this
The surface tension of pipeline is able to through this microchannel.
Capillary valves are to carry out hydrophilic treated to microchannel, and when liquid does not infiltrate this pipeline, liquid is under the action of the centrifugal force
Bending lower section is rested on, liquid infiltrates entire microchannel when centrifugal force stops, then implements centrifugal force, and liquid is in centrifugal force and hair
It, being capable of drive to the basket this microchannel under the action of thin power.
Preferably, second mixed cell is set between the 5th valve and the second reaction member.
Preferably, second mixed cell includes the second mixing channel.
Preferably, second mixing channel is arc-shaped or rectangular etc., those skilled in the art can according to actual needs into
Row selection.
Preferably, the biochemistry detection area also sets up second liquid storage unit, and second liquid storage unit passes through micro- logical
Road is connected to the second mixed cell；The second liquid storage unit includes fluid storage tanks, and the fluid storage tanks include storage
Flow container tank body, the inside of the liquid storage tank are equipped with the cavity for accommodating liquid reagent；First sealing-plug, the first sealing-plug and institute
It states liquid storage tank to be detachably connected, and one end of the cavity can be sealed；The other end of cavity can be by micro- logical
Road is connected to the first reaction member and/or the second allocation unit；And cavity connects with the first reaction member and/or the second allocation unit
Hot melt sealing-plug is set on logical microchannel.When needing to discharge liquid, using laser or the mode of heating by fluid storage tanks
Hot melt sealing-plug fusing.The liquid of the fluid storage tanks storage in the immune detection area can be dilution property liquid, bottom
Thing liquid and cleaning solution etc..
Preferably, the biochemistry detection area also sets up third allocation unit, and sample injection unit passes through third allocation unit and the
The connection of two reaction members.
Preferably, the third allocation unit is set between the first valve and the second reaction member.When biochemistry detection area is set
When setting multiple second reaction members, the third allocation unit is used to distribute prepare liquid to each second reaction member.
Preferably, the third allocation unit includes third distributing trough.
Preferably, the third distributing trough is arc-shaped or rectangular etc., the shape art technology of the third distributing trough
Personnel can select according to actual needs.
Preferably, the biochemistry detection area includes multiple second reaction members, such as 2,3,5,8,10,15
A, 18,25 or 30 etc., the multiple second reaction member is connected to third allocation unit.In the present invention, such as without spy
Different explanation, it is described " multiple " to refer at least two.
Preferably, the second curing agent layer choosing is from biochemical reagents cured layer.
Preferably, the sample injection unit, the 4th quantitative slot and the second reaction member apart from substrate centrifugation center distance according to
Secondary increase.
Preferably, fixing groove is arranged in the geometric center of the substrate.The substrate can be to be round or rectangular etc., and the application is not
Its shape is defined.
Preferably, the substrate is transparent substrates, preferably in PMMA substrate, PS substrate, COP substrate or COC substrate
Any one or at least two combination, typical but non-limiting combination such as PMMA substrate and PS substrate, COP substrate and COC
Substrate, PMMA substrate, PS substrate and COP substrate.
The micro-fluidic chip based on immune detection and biochemistry detection includes substrate, base as a preferred technical solution,
Sample injection unit, immune detection area and biochemistry detection area is arranged in plate；
Sample injection unit includes sample introduction tank, the first quantitative slot, collecting tank and the first distributing trough, sample introduction tank, the first quantitative slot and
Collecting tank passes sequentially through microchannel connection, and the first quantitative slot is also connected to the first distributing trough；First quantitative slot and the first distributing trough
First valve is set on the microchannel of connection；
Immune detection area includes the second quantitative slot being sequentially communicated, the second allocation unit, at least one first reaction member
With devil liquor recovery unit；Further include the first liquid storage unit, solidification oxidant layer is provided in the first reaction member；
Second allocation unit includes the second distributing trough；First reaction member includes third quantitative slot, the first mixing channel and
One reactive tank；First liquid storage unit includes fluid storage tanks；Devil liquor recovery unit includes waste liquid tank；
Second distributing trough, third quantitative slot, the first mixing channel, the first reactive tank and waste liquid tank, wherein the second quantitative slot with
The connection of first distributing trough；Fluid storage tanks are connected to the second distributing trough and/or third quantitative slot；Second quantitative slot and the second distribution
Second valve is set on the microchannel of slot connection, the 4th valve is set on the microchannel that third quantitative slot is connected to the first mixing channel, the
Third valve is set on the microchannel that one reactive tank is connected to waste liquid tank；
Biochemistry detection area includes that the 4th quantitative slot being sequentially communicated, the second mixed cell, third allocation unit and second are anti-
Answer unit；It further include second liquid storage unit, second liquid storage unit is connected to the second mixed cell, the second reaction member
In be provided with the second solidification oxidant layer；
Second mixed cell includes the second mixing channel, and third allocation unit includes third distributing trough, the second reaction member packet
Include the second reactive tank；Second liquid storage unit includes fluid storage tanks；
4th quantitative slot is connected to the first distributing trough, and is arranged on the microchannel that the 4th quantitative slot is connected to the second mixing channel
The 6th valve is arranged on the microchannel that the second mixing channel is connected to third distributing trough for five valves.
In the present invention, first quantitative slot can be used for the total serum amount that this quantitative chip needs, and the 4th quantitative slot can be used for
The total serum amount of quantitative biochemical detection, the second quantitative slot can be used for the total serum amount of quantitative immunological detection, and third quantitative slot is available
In the serum amount or substrate solution or dilution etc. of quantitatively single immune detection project.
The second object of the present invention is to provide a kind of detector based on immune detection and biochemistry detection, the detector
Including micro-fluidic chip as described above.
Preferably, the detector based on immune detection and biochemistry detection further include sensing chamber, centrifugal device, light source,
Light signal receiving, light signal processing device and display device；The substrate is fixed in centrifugal device, and is located at sensing chamber
In；Light source is set in sensing chamber, is connected with display device；Light signal receiving is set in sensing chamber, passes through optical signal
Processing unit is connected with display device.
Preferably, the detector based on immune detection and biochemistry detection further includes optical signal enhancement device, preferably
Photomultiplier tube, light signal receiving are connected by optical signal enhancement device with light signal processing device.
Preferably, the light signal receiving includes light-receiving probe.
It is described that large size detection device --- POCT (care diagnostic) is compared based on the detector of immune detection and biochemistry detection
Product has cost advantage, and equipment simple operations are simple；It compares existing POCT product: having the production of individually detection biochemical indicator at present
Product increase immune detection index, demand are immunized simultaneously the providing convenience property of demand with biochemical indicator, more efficiently.
It include as a preferred technical solution, as follows using the method that above-mentioned detector carries out immune detection and biochemistry detection
Step:
(1) a certain amount of whole blood is added into sample introduction tank, is put into centrifugal device, chip starts to rotate；
(2) with rotation, whole blood enters in the first quantitative slot and collecting tank, and after a period of time, erythrocyte is due to centrifugation
Power sinks to collecting tank, blood plasma or serum keeping in the first quantitative slot；
(3) micro-fluidic chip is rotated, the blood plasma or serum in the first quantitative slot break through the first valve, are assigned to the 4th quantitative slot
In this second quantitative slot；
(4) by Fixed Point Laser or the method for heating, the liquid storage in immune detection area at third quantitative slot is opened
The valve of fluid storage tanks in tank and biochemistry detection area, rotary chip (counterclockwise) discharge dilution, dilution are discharged completely
To third quantitative slot and the second mixing channel；
(5) rotate micro-fluidic chip, by the 4th quantitative slot in the second quantitative slot blood plasma or serum be discharged into second and mix
Slot and third quantitative slot；
(6) micro-fluidic chip is rotated, liquid enters in the second reactive tank in the second mixing channel, dissolves in the second reactive tank
Solidified reagents, and reacted, complete biochemical reaction；Liquid in third quantitative slot enters the first mixing channel, dissolution first
Cured enzyme labelled antibody in mixing channel enters the first reactive tank after being sufficiently mixed；
(7) later period is rotated, biochemical test window is automatically brought to, carries out biochemistry detection；Fixed antibody in first reactive tank
It is combined with antigen in serum, forms fixed antibody-antigene-enzyme labelled antibody interlayer structure onboard；
(8) after completing biochemistry detection, by Fixed Point Laser or the method for heating, liquid storage at the second distributing trough is opened
The valve of tank discharges cleaning solution, rotates micro-fluidic chip, wash liquid stream is made to be cleaned, be washed through the further channel of immune detection
Unbonded enzyme labelled antibody is removed, free antigen finally drains the liquid in the first reactive tank；
(9) by Fixed Point Laser or the method for heating, the valve of fluid storage tanks at the second distributing trough is opened, is rotated micro-
Fluidic chip discharges substrate solution, carries out chemiluminescence immunoassay reaction, and chip is gone under chemiluminescent assay window, after a few minutes,
It is reacted into plateau, carries out test luminous signal.
The revolving speed of the rotating centrifugal can be 1000-8000r/min, as 1500r/min, 2000r/min, 2300r/min,
2800r/min, 3000r/min, 5000r/min, 7000r/min or 7500r/min etc., the time of rotation can be 1-10min, such as
2min, 3min, 5min, 8min, 9min or 9.5min etc..
The third object of the present invention is to provide a kind of immune detection and biochemical detection methods, and the method includes walking as follows
It is rapid:
(1) sample to be tested is added, is centrifuged by rotation micro-fluidic chip so that sample to be tested flows；
(2) rotation micro-fluidic chip makes sample to be tested flow into immune detection area and biochemistry detection area, is immunized respectively
Reaction and biochemical reaction；
(3) rotation micro-fluidic chip makes the liquid discharge after immune response.
Chemical luminous immune detection method provided by the invention is based on centrifugation technique, by the rotation for controlling micro-fluidic chip
The position that sample flows into is controlled, and then completes chemiluminescence immunoassay detection.
The revolving speed of the rotation can be 1000-8000r/min, as 1500r/min, 2000r/min, 2300r/min,
2800r/min, 3000r/min, 5000r/min, 7000r/min or 7500r/min etc., the time of rotation can be 1-10min, such as
2min, 3min, 5min, 8min, 9min or 9.5min etc..
Numberical range of the present invention not only includes enumerated point value, further includes the above-mentioned numerical value not included
Arbitrary point value between range, as space is limited and for concise consideration, range described in the present invention no longer exclusive list includes
Specific point value.
Compared with prior art, the invention has the benefit that
1, the detector provided by the invention based on immune detection and biochemistry detection has been compatible with immune detection and biochemistry detection,
Two distinct types of detection is integrated on a single die, by realizing two different detections, Neng Goubang on an instrument
Doctor is helped to be quickly obtained as a result, helping illness analysis；
2, the detector structure provided by the invention based on immune detection and biochemistry detection is simple, using dual probe mode,
Independent detection is independent of each other；
3, finger tip blood or vein can be directly used in the detector provided by the invention based on immune detection and biochemistry detection
Other position whole bloods such as blood, are applied directly on micro-fluidic chip, realize blood plasma separation and quantitative by centrifugal separation equipment, are assigned to
Instrument connection；Whole process is easy to operate, is not necessarily to professional person, without blood plasma is got out in advance, tester only needs to understand fetching
Sharp blood；
4, the detector scalability provided by the invention based on immune detection and biochemistry detection is strong, can equipped with bluetooth and
WiFi interface, facilitates and transmits data and other equipment.
Detailed description of the invention
Fig. 1 is the structure connection diagram for the detector based on immune detection and biochemistry detection that embodiment 3 provides.
Fig. 2 is that the structure of the micro-fluidic chip for the detector based on immune detection and biochemistry detection that embodiment 3 provides is shown
It is intended to.
Fig. 3 is the front view for the fluid storage tanks that embodiment 3 provides.
Fig. 4 is the top view for the fluid storage tanks that embodiment 3 provides.
Wherein, 1, sample introduction tank；2, the first quantitative slot；3, collecting tank；4, the first valve；5, the 4th quantitative slot；6, the first dilution
Liquid storage tank；7, the second mixing channel；8, the second reactive tank；9, fixing groove；10, the second quantitative slot；11, substrate solution storage tank；12,
Cleaning solution storage tank；13, the second dilution holding vessel；14, third quantitative slot；15, the first mixing channel；16, the first reactive tank；
17, waste liquid tank；18, fluid storage tanks；19, hot melt sealing-plug；20, microchannel；21, board-on-chip；22, photomultiplier tube；
23, immune detection hole；24, it is emitted light probe；25, centrifuge；26, optical fiber；27, light source；28, display；29, spectrometer；30,
Incident light probe；31, biochemical reaction hole.
Specific embodiment
To further illustrate the technical scheme of the present invention below with reference to the accompanying drawings and specific embodiments.
Embodiment 1
A kind of detector based on immune detection and biochemistry detection, including micro-fluidic chip, the micro-fluidic chip include
Sample injection unit, immune detection area and biochemistry detection area are arranged on substrate for substrate；
Sample injection unit includes sample introduction tank；
Immune detection area includes the first reaction member and devil liquor recovery unit, and the first reaction member and devil liquor recovery unit are logical
Cross the microchannel connection for being provided with third valve；First reaction member includes the first reactive tank, and the is fixed in the first reaction member
One solidification oxidant layer；Devil liquor recovery unit includes waste liquid tank；
Biochemistry detection area includes the second reaction member, and the second reaction member includes the second reactive tank, in the second reaction member
It is fixed with the second solidification oxidant layer；
Sample injection unit is connected to the first reaction member and the second reaction member respectively by being provided with the microchannel of the first valve.
The sample injection unit, the second quantitative slot, the first reaction member and devil liquor recovery cell distance substrate are centrifuged neutral
Distance successively increases；
The distance of the sample injection unit, the 4th quantitative slot and the second reaction member apart from substrate centrifugation center successively increases；
The detector based on immune detection and biochemistry detection further includes that sensing chamber, centrifugal device, light source, optical signal connect
Receiving apparatus, light signal processing device, optical signal enhancement device and display device；The substrate is fixed in centrifugal device, and position
In sensing chamber；Light source is set in sensing chamber, is connected with display device；Light signal receiving is set in sensing chamber, is led to
It crosses optical signal enhancement device and light signal processing device is connected with display device；
The optical signal enhancement device is photomultiplier tube；The light signal receiving includes light-receiving probe, preferably
It is respectively used to receive the optical signal in immune detection area and the optical signal in biochemistry detection area for 2.Light signal processing device includes light
Spectrometer.
Included the following steps: using the method that above-mentioned preferred detector carries out immune detection and biochemistry detection
(1) testing liquid is added into sample introduction tank, by rotating micro-fluidic chip sample to be tested is flowed；
(2) micro-fluidic chip is rotated, sample to be tested flows into the first reaction member and the second reaction member by the first valve, point
Immune response and biochemical reaction are not carried out with the first solidified reagents and the second solidified reagents；Capture optical signal；
(3) rotation micro-fluidic chip makes the liquid discharge after immune response.
Wherein, when biochemistry detection, incident light light probe is passed to through optical fiber by the light that light source issues, is irradiated to biochemical reaction hole
On, signal is received by outgoing light probe, in feedback to spectrometer, by inter-process, as the result is shown over the display.It is immune
When detection, the light that immune detection hole issues is received by photomultiplier tube, in feedback to spectrometer, is analyzed, as the result is shown
Over the display.
The revolving speed of the rotating centrifugal can be 1000-8000r/min, as 1500r/min, 2000r/min, 2300r/min,
2800r/min, 3000r/min, 5000r/min, 7000r/min or 7500r/min etc., the time of rotation can be 1-10min, such as
2min, 3min, 5min, 8min, 9min or 9.5min etc..
Embodiment 2
A kind of detector based on immune detection and biochemistry detection includes micro-fluidic chip, and the micro-fluidic chip includes base
Sample injection unit, immune detection area and biochemistry detection area is arranged in plate, substrate；
Sample injection unit includes sample introduction tank, the first quantitative slot, collecting tank and the first distributing trough, sample introduction tank, the first quantitative slot and
Collecting tank passes sequentially through microchannel connection, and the first quantitative slot is also connected to the first distributing trough；First quantitative slot and the first distributing trough
First valve is set on the microchannel of connection；
Immune detection area includes the second quantitative slot being sequentially communicated, the second allocation unit, at least one first reaction member
With devil liquor recovery unit；Further include the first liquid storage unit, solidification oxidant layer is provided in the first reaction member；
Second allocation unit includes the second distributing trough；First reaction member includes third quantitative slot, the first mixing channel and
One reactive tank；First liquid storage unit includes cleaning solution storage tank and substrate solution storage tank；Devil liquor recovery unit includes waste liquid
Leakage fluid dram is arranged in slot, waste liquid tank；
Second distributing trough, third quantitative slot, the first mixing channel, the first reactive tank and waste liquid tank, wherein the second quantitative slot with
The connection of first distributing trough；Cleaning solution storage tank and substrate solution storage tank are connected to the second distributing trough；Second quantitative slot and second
Second valve is set on the microchannel of distributing trough connection, is arranged the 4th on the microchannel that third quantitative slot is connected to the first mixing channel
Third valve is arranged on the microchannel that the first reactive tank is connected to waste liquid tank for valve；
Biochemistry detection area includes that the 4th quantitative slot being sequentially communicated, the second mixed cell, third allocation unit and second are anti-
Answer unit；The second solidification oxidant layer is provided in second reaction member；
Second mixed cell includes the second mixing channel, and third allocation unit includes third distributing trough, the second reaction member packet
Include the second reactive tank；
4th quantitative slot is connected to the first distributing trough, and is arranged on the microchannel that the 4th quantitative slot is connected to the second mixing channel
The 6th valve is arranged on the microchannel that the second mixing channel is connected to third distributing trough for five valves；
First valve, the second valve, third valve, the 4th valve, the 5th valve and the 6th valve are drain valve or capillary valves.
The sample injection unit, the second quantitative slot, the first reaction member and devil liquor recovery cell distance substrate are centrifuged neutral
Distance successively increases；
The distance of the sample injection unit, the 4th quantitative slot and the second reaction member apart from substrate centrifugation center successively increases；
The detector based on immune detection and biochemistry detection further includes that sensing chamber, centrifugal device, light source, optical signal connect
Receiving apparatus, light signal processing device, optical signal enhancement device and display device；The substrate is fixed in centrifugal device, and position
In sensing chamber；Light source is set in sensing chamber, is connected with display device；Light signal receiving is set in sensing chamber, is led to
It crosses optical signal enhancement device and light signal processing device is connected with display device.
The optical signal enhancement device is photomultiplier tube；The light signal receiving includes light-receiving probe, preferably
It is respectively used to receive the optical signal in immune detection area and the optical signal in biochemistry detection area for 2；Light signal processing device includes light
Spectrometer.
Included the following steps: using the method that above-mentioned preferred detector carries out immune detection and biochemistry detection
(1) a certain amount of whole blood is added into sample introduction tank, is put into centrifugal device, chip starts to rotate；
(2) with rotation, whole blood enters in the first quantitative slot and collecting tank, and after a period of time, erythrocyte is due to centrifugation
Power sinks to collecting tank, blood plasma or serum keeping in the first quantitative slot；
(3) micro-fluidic chip is rotated, the blood plasma or serum in the first quantitative slot break through the first valve, are assigned to the 4th quantitative slot
In the second quantitative slot；
(5) rotation micro-fluidic chip by the 4th quantitative slot and the second quantitative slot blood plasma or serum be discharged into the second mixing channel
With third quantitative slot；
(6) micro-fluidic chip is rotated, liquid enters in the second reactive tank in the second mixing channel, dissolves in the second reactive tank
Solidified reagents, and reacted, complete biochemical reaction；Liquid in third quantitative slot enters the first mixing channel, dissolution first
Cured enzyme labelled antibody in mixing channel enters the first reactive tank after being sufficiently mixed；
(7) later period is rotated, biochemical test window is automatically brought to, carries out biochemistry detection；Fixed antibody in first reactive tank
It is combined with antigen in serum, forms fixed antibody-antigene-enzyme labelled antibody interlayer structure onboard；
(8) after completing biochemistry detection, by Fixed Point Laser or the method for heating, liquid storage at the second distributing trough is opened
The valve of tank, discharges cleaning solution, and rotation micro-fluidic chip makes wash liquid stream be cleaned, be washed through the further channel of immune detection
Unbonded enzyme labelled antibody is removed, free antigen finally drains the liquid in the first reactive tank；
(9) by Fixed Point Laser or the method for heating, the valve of fluid storage tanks at the second distributing trough is opened, is rotated micro-
Fluidic chip discharges substrate solution, carries out chemiluminescence immunoassay reaction, and chip is gone under chemiluminescent assay window, after a few minutes,
It is reacted into plateau, carries out test luminous signal.
Wherein, when biochemistry detection, incident light light probe is passed to through optical fiber by the light that light source issues, is irradiated to biochemical reaction hole
On, signal is received by outgoing light probe, in feedback to spectrometer, by inter-process, as the result is shown over the display.It is immune
When detection, the light that immune detection hole issues is received by photomultiplier tube, in feedback to spectrometer, is analyzed, as the result is shown
Over the display.
The revolving speed of the rotating centrifugal can be 1000-8000r/min, as 1500r/min, 2000r/min, 2300r/min,
2800r/min, 3000r/min, 5000r/min, 7000r/min or 7500r/min etc., the time of rotation can be 1-10min, such as
2min, 3min, 5min, 8min, 9min or 9.5min etc..
Embodiment 3
A kind of detector (as shown in Figure 1) based on immune detection and biochemistry detection, including micro-fluidic chip is (such as Fig. 2 institute
Show), the micro-fluidic chip includes substrate, and sample injection unit, immune detection area and biochemistry detection area is arranged in substrate；
Sample injection unit includes sample introduction tank 1, the first quantitative slot 2, collecting tank 3 and the first distributing trough, and sample introduction tank 1, first is quantitative
Slot 2 passes sequentially through microchannel with collecting tank 3 and is connected to, and the first quantitative slot 2 is also connected to the first distributing trough；First quantitative slot 2 and
The first valve 4 is set on the microchannel of one distributing trough connection；
Immune detection area includes the second quantitative slot 10 being sequentially communicated, the second allocation unit, at least one first reaction list
Member and devil liquor recovery unit；Further include the first liquid storage unit, solidification oxidant layer is provided in the first reaction member；
Second allocation unit includes the second distributing trough；First reaction member includes third quantitative slot 14, the first mixing channel 15
(there is cured enzyme labelled antibody in its front end) and the first reactive tank 16 (fixed antibody is arranged at its bottom)；First liquid storage unit packet
Include the second dilution storage tank 13, cleaning solution storage tank 12 and substrate solution storage tank 11 (the second dilution storage tank 13, cleaning solution
The structure of storage tank 12 and substrate solution storage tank 11 is identical as the structure of fluid storage tanks 18, such as Fig. 3 and Fig. 4), devil liquor recovery
Unit includes waste liquid tank 17；
Second distributing trough, third quantitative slot 14, the first mixing channel 15, the first reactive tank 16 and waste liquid tank 17, wherein second
Quantitative slot 10 is connected to the first distributing trough；Second dilution storage tank 13 is connected to third quantitative slot 14, cleaning solution storage tank 12
It is connected to substrate solution storage tank 11 with the second distributing trough；Is arranged on the microchannel that second quantitative slot 10 is connected to the second distributing trough
The 4th valve, the first reactive tank 16 and waste liquid tank are arranged on the microchannel that third quantitative slot 14 is connected to the first mixing channel 15 for two valves
Third valve is set on the microchannel of 17 connections；
Biochemistry detection area includes that the 4th quantitative slot 5 being sequentially communicated, the second mixed cell, third allocation unit and second are anti-
Answer unit；It further include second liquid storage unit, second liquid storage unit is connected to the second mixed cell, the second reaction member
In be provided with the second solidification oxidant layer；
Second mixed cell includes the second mixing channel 7, and third allocation unit includes third distributing trough, the second reaction member packet
Include the second reactive tank 8 (there are cured biochemical reagents inside it)；Second liquid storage unit includes the first dilution storage tank 6；
First dilution storage tank 6, the second dilution storage tank 13, substrate solution storage tank 11 and cleaning solution storage tank 12
The structure (as shown in Figure 3 and Figure 4) of structure such as fluid storage tanks 18, the fluid storage tanks 18 include liquid storage tank, described
The inside of liquid storage tank is equipped with the cavity for accommodating liquid reagent；First sealing-plug, the first sealing-plug and the liquid storage tank
It is detachably connected, and one end of the cavity can be sealed；The other end of cavity can be anti-by microchannel 20 and first
Unit and/or the second allocation unit is answered to be connected to；And the microchannel that cavity is connected to the first reaction member and/or the second allocation unit
Hot melt sealing-plug 19 is set on 20.When needing to discharge liquid, using laser or the mode of heating by the hot melt of fluid storage tanks
Property sealing-plug 19 melt.The microchannel is set to the board-on-chip 21 of micro-fluidic chip.
4th quantitative slot 5 is connected to the first distributing trough, is set on the microchannel that the 4th quantitative slot 5 is connected to the second mixing channel 7
The 5th valve is set, the 6th valve is set on the microchannel that the second mixing channel 7 is connected to third distributing trough；
First valve 4, the second valve, third valve, the 4th valve, the 5th valve and the 6th valve drain valve or capillary valves.
The sample injection unit, the second quantitative slot 10, the first reaction member and the centrifugation of devil liquor recovery cell distance substrate are neutral
Distance successively increase；
The distance of the sample injection unit, the 4th quantitative slot 5 and the second reaction member apart from substrate centrifugation center successively increases；
The substrate is provided with the fixing groove 9 being fixed on centrifuge 25.
The detector based on immune detection and biochemistry detection further includes sensing chamber, centrifugal device (centrifuge 25), light
Source 27, light signal receiving, light signal processing device, optical signal enhancement device and display device；The substrate be fixed on from
In center device, and it is located in sensing chamber；Light source 27 is set in sensing chamber, is connected with display device；Light signal receiving is set
It is placed in sensing chamber, is connected by optical signal enhancement device and light signal processing device with display device.
The optical signal enhancement device is photomultiplier tube 22；The light signal receiving includes that light-receiving probe (enters
Penetrate light probe 30 and outgoing light probe 24), preferably 2 optical signal and the biochemistry detection areas for being respectively used to receive immune detection area
Optical signal；Light signal processing device includes spectrometer 29.
Included the following steps: using the method that above-mentioned preferred detector carries out immune detection and biochemistry detection
(1) a certain amount of whole blood is added into sample introduction tank 1, is put into centrifugal device, chip starts to rotate；
(2) with rotation, whole blood enters in the first quantitative slot 2 and collecting tank 3, after a period of time, erythrocyte due to from
Mental and physical efforts sink to collecting tank 3, blood plasma or serum keeping in the first quantitative slot 2；
(3) turn rotation, and the blood plasma or serum in the first quantitative slot 2 break through the first valve 4, is assigned to the 4th quantitative slot
In 5 this second quantitative slot 10；
(4) it by Fixed Point Laser or the method for heating, so that hot melt sealing-plug 19 melts, opens in immune detection area
The valve of first dilution storage tank 6 in the second dilution storage tank 13 and biochemistry detection area at third quantitative slot 14, rotation
Chip discharges dilution, dilution is completely discharged to third quantitative slot 14 and the second mixing channel 7；
(5) turn rotation, by the 4th quantitative slot 5 in the second quantitative slot 10 blood plasma or serum be discharged into second and mix
Slot 7 and third quantitative slot 14；
(6) turn rotation, and liquid enters in the second reactive tank 8 in the second mixing channel 7, dissolves in the second reactive tank 8
Solidified reagents, and reacted, complete biochemical reaction；Liquid in third quantitative slot 14 enters the first mixing channel 15, molten
Cured enzyme labelled antibody in the first mixing channel 15 is solved, the first reactive tank 16 is entered after being sufficiently mixed；
(7) later period is rotated, biochemical test window is automatically brought to, carries out biochemistry detection；It is fixed in first reactive tank 16 to resist
Body is combined with antigen in serum, forms fixed antibody-antigene-enzyme labelled antibody interlayer structure onboard；
(8) it after completing biochemistry detection, by Fixed Point Laser or the method for heating, opens cleaning solution at the second distributing trough and deposits
The valve of storage tank 12, discharges cleaning solution, and rotation makes wash liquid stream be cleaned through the further channel of immune detection, wash away and do not tie
The enzyme labelled antibody of conjunction, free antigen finally drain the liquid in the first reactive tank 16；
(9) by Fixed Point Laser or the method for heating, the valve of substrate solution storage tank 11 at the second distributing trough, rotation are opened
Turn, discharge substrate solution, carry out chemiluminescence immunoassay reaction, chip is gone under chemiluminescent assay window, after a few minutes, react into
Enter plateau, carries out test luminous signal.
Wherein, when biochemistry detection, the light issued by light source 27 passes to incident light light probe through optical fiber 26, is irradiated to biochemical anti-
It answers on hole 31, signal is received by outgoing light probe 24, in feedback to spectrometer 29, by inter-process, as the result is shown aobvious
Show on device 28.When immune detection, the light that immune detection hole 23 issues is received by photomultiplier tube 22, and feedback arrives spectrometer 29
On, it is analyzed, as the result is shown on the display 28.
The revolving speed of the rotating centrifugal can be 1000-8000r/min, as 1500r/min, 2000r/min, 2300r/min,
2800r/min, 3000r/min, 5000r/min, 7000r/min or 7500r/min etc., the time of rotation can be 1-10min, such as
2min, 3min, 5min, 8min, 9min or 9.5min etc..
Application Example
Immune detection and biochemistry detection are carried out based on the detector of immune detection and biochemistry detection using described in embodiment 3
Enzymic-labelled antibody process: the AFP antibody of alkali phosphatase enzyme mark is using the enzyme mark in 104198721 B of patent CN
Note method.It is accurate to measure alkaline phosphatase 0.5-0.8mg, it is placed in the centrifuge tube of 2mL；Weigh the SMCC (Malaysia the N- acyl of 4-6mg
Formimino group) hexamethylene -1- carboxylic acid succinimide ester with DMSO (dimethyl sulfoxide) is dissolved to a certain concentration；Containing alkalinity
Suitable SMCC solution is added in the centrifuge tube of phosphatase, after mixing, is reacted 1 hour in 25 DEG C, after, dialysis removal solution
In excessive SMCC, obtain the alkaline phosphatase of coupled antibody using AKTA protein purification instrument.
Enzyme labelled antibody solidification process: enzyme labelled antibody after purification is concentrated with super filter tube, is re-dissolved in freeze-drying stabilizer, this
Freeze-drying stabilizer include sucrose, histidine buffering liquid (pH=7.4), polysorbate -20 mixed solution, be placed in freeze-drying
It is lyophilized 12-24 hours in machine, is dispensed in kit at dry powder-shaped loaded in brown vial.
The fixed onboard process of antibody:
Through surface hydrophilic modification and rich in-NH at micro-fluidic chip coated antibody2Group, content is 0.21eq/g,
Micro-fluidic chip is immersed in PBS (pH=8.0) container of 100mL0.01M first, addition 200uL glutaraldehyde solution, then plus
Entering the AFP coated antibody of 300ug, slight concussion mixes mixed solution, and it is protected from light and is placed in 4 DEG C of refrigerators, after reaction 12 hours,
Chip is taken out, closes 2 at 4 DEG C with Tris (pH=7.4) confining liquid of the TWEEN-20 of the BSA containing 0.5% and 0.01%
Hour, micro-fluidic chip is cleaned 4-6 times with the PBS of 10mM finally, is dried 2 hours in 50 DEG C of baking oven, obtains certain area
Domain is coated with the micro-fluidic chip of AFP primary antibody.
Biochemical reagents solidification process:
It prepares biochemistry detection reagent: accurately weighing 1.50g sodium dihydrogen phosphate, 0.30g disodium ethylene diamine tetraacetate, 0.40g
Kathon, 0.05g4- hydroxyethyl piperazine propane sulfonic acid, 0.04g ninhydrin, 1.25g sodium citrate, 0.50g glycerol, it is water-soluble with purifying
Solution, is configured to the solution of 100ml, as biochemistry detection reagent.
Biochemical reagents freeze-drying: it is spare that biochemistry detection reagent solution is frozen into dry powder in vacuum freezing drying oven.
Powdered reagent sample-adding: the dry biochemical inspection of quality needed for being added in chip biochemical reaction hole in chip fabrication process
Survey powder.
Enzyme-catalyzed chemical luminescence detection process:
By the serum blending incubation of enzyme labelled antibody and people to be measured, the region incubation reaction certain time of coating primary antibody is flowed into
Afterwards, Tris (pH=7.4) cleaning solution 100uL of the Trixon-100 containing 0.01% is injected, after cleaning 3 times, reinjects 190uL
Substrate A MPPD and its adamantane derivative CSPD and CDP-Star.
Enzymatic makes luminous substrate continued for constant shine, with the relative light intensity of luminous tester measurement reaction product, then root
The result of determinand is obtained according to the concentration of calibration object.
The Applicant declares that the foregoing is merely a specific embodiment of the invention, but protection scope of the present invention not office
It is limited to this, it should be clear to those skilled in the art, any to belong to those skilled in the art and take off in the present invention
In the technical scope of dew, any changes or substitutions that can be easily thought of, and all of which fall within the scope of protection and disclosure of the present invention.
Claims (10)
1. a kind of micro-fluidic chip based on immune detection and biochemistry detection, which is characterized in that the micro-fluidic chip includes base
Sample injection unit, immune detection area and biochemistry detection area are arranged on substrate for plate；
Sample injection unit includes sample introduction tank；
Immune detection area includes the first reaction member and devil liquor recovery unit, and the first reaction member and devil liquor recovery unit pass through micro-
Channel connection；First reaction member includes the first reactive tank, is fixed with the first solidification oxidant layer in the first reaction member；Devil liquor recovery
Unit includes waste liquid tank；
Biochemistry detection area includes the second reaction member, and the second reaction member includes the second reactive tank, is fixed in the second reaction member
There is the second solidification oxidant layer；
Sample injection unit is connected to the first reaction member and the second reaction member respectively by microchannel.
2. the micro-fluidic chip according to claim 1 based on immune detection and biochemistry detection, which is characterized in that it is described into
Sample unit is connected to the first reaction member and the second reaction member respectively by being provided with the microchannel of the first valve；
Preferably, first reaction member is connected to devil liquor recovery unit by being provided with the microchannel of third valve；
Preferably, the sample injection unit further includes the first quantitative slot and collecting tank, and sample introduction tank, the first quantitative slot and collecting tank are successively
It is connected to by microchannel；
Preferably, first quantitative slot reacts single by being equipped with the microchannel of the first valve with the first reaction member and second respectively
Member connection；
Preferably, the sample injection unit further includes the first distributing trough, and the first quantitative slot is anti-with first respectively by the first distributing trough
Unit and the second reaction member is answered to be connected to；
Preferably, the first distributing trough is set between the first valve and the first reaction member or the second reaction member.
3. the micro-fluidic chip according to claim 1 or 2 based on immune detection and biochemistry detection, which is characterized in that institute
It states immune detection area and also sets up the second quantitative slot, and the second quantitative slot passes through microchannel respectively and reacts single with sample injection unit and first
Member connection；
Preferably, the second quantitative slot is set between the first valve and the first reaction member；
Preferably, the second valve is set on the microchannel that the second quantitative slot is connected to the first reaction member；
Preferably, second quantitative slot is set between the first distributing trough and the first reaction member；
Preferably, the immune detection area also sets up the second allocation unit, the second allocation unit respectively with the second quantitative slot and
The connection of one reactive tank；
Preferably, the second allocation unit is set between the second valve and the first reaction member；
Preferably, second allocation unit includes the second distributing trough.
4. being according to claim 1 the micro-fluidic chip based on immune detection and biochemistry detection described in one of -3, feature exists
In the immune detection area further includes the first liquid storage unit, the first liquid storage unit and the first reaction member and/or
Two allocation units are connected to by microchannel, are preferably connected to the second allocation unit by microchannel；
Preferably, first liquid storage unit includes at least one fluid storage tanks, and first fluid storage tanks include
Liquid storage tank, the inside of the liquid storage tank are equipped with the cavity for accommodating liquid reagent；First sealing-plug, the first sealing-plug with
The liquid storage tank is detachably connected, and can seal one end of the cavity；The other end of cavity can be by micro-
Channel is connected to the first reaction member and/or the second allocation unit；And cavity and the first reaction member and/or the second allocation unit
Hot melt sealing-plug is set on the microchannel of connection.
5. the micro-fluidic chip described in one of -4 based on immune detection and biochemistry detection according to claim 1, which is characterized in that
First reaction member further includes third quantitative slot and the first mixing channel, and sample to be tested can be successively by the by microchannel
Three quantitative slots, the first mixing channel and the first reactive tank；
Preferably, the 4th valve is set on the microchannel that third quantitative slot is connected to the first mixing channel；
Preferably, the third quantitative slot is also connected to liquid storage unit by microchannel；
Preferably, the immune detection area includes multiple first reaction members, and the multiple first reaction member is respectively with second
Allocation unit connection；
Preferably, the first curing agent layer choosing self-curing reagent layer；
Preferably, the sample injection unit, the second quantitative slot, the first reaction member and the centrifugation of devil liquor recovery cell distance substrate are neutral
Distance successively increase.
6. the micro-fluidic chip described in one of -5 based on immune detection and biochemistry detection according to claim 1, which is characterized in that
The biochemistry detection area also sets up the 4th quantitative slot, and sample injection unit is connected to by the 4th quantitative slot with the second reaction member；
Preferably, the 4th quantitative slot is set between the first valve and the second reaction member, and the 4th quantitative slot is reacted with second
The 5th valve is set on the microchannel of unit connection；
Preferably, the 4th quantitative slot is connected to the first distributing trough.
7. the micro-fluidic chip described in one of -6 based on immune detection and biochemistry detection according to claim 1, which is characterized in that
The biochemistry detection area also sets up the second mixed cell, and sample injection unit is connected to by the second mixed cell with the second reaction member,
Second mixed cell is set between the first valve and the second reaction member；
Preferably, the 6th valve is set on the microchannel that second mixed cell is connected to the second reaction member；
Preferably, first valve, the second valve, third valve, the 4th valve, the 5th valve and the 6th valve independently selected from drain valve and/
Or capillary valves；
Preferably, second mixed cell is set between the 5th valve and the second reaction member；
Preferably, second mixed cell includes the second mixing channel；
Preferably, the biochemistry detection area also sets up second liquid storage unit, second liquid storage unit by microchannel with
The connection of second mixed cell；The second liquid storage unit includes fluid storage tanks, and the fluid storage tanks include fluid reservoir
Tank body, the inside of the liquid storage tank are equipped with the cavity for accommodating liquid reagent；First sealing-plug, the first sealing-plug and the storage
Flow container tank body is detachably connected, and can seal one end of the cavity；The other end of cavity can by microchannel with
First reaction member and/or the connection of the second allocation unit；And cavity is connected to the first reaction member and/or the second allocation unit
Hot melt sealing-plug is set on microchannel；
Preferably, the biochemistry detection area also sets up third allocation unit, and sample injection unit is anti-with second by third allocation unit
Unit is answered to be connected to；
Preferably, the third allocation unit is set between the first valve and the second reaction member；
Preferably, the third allocation unit includes third distributing trough；
Preferably, the biochemistry detection area includes multiple second reaction members, and the multiple second reaction member divides with third
It is connected to unit；
Preferably, the second curing agent layer choosing is from biochemical reagents cured layer；
Preferably, the distance of the sample injection unit, the 4th quantitative slot and the second reaction member apart from substrate centrifugation center successively increases
Add；
Preferably, fixing groove is arranged in the geometric center of the substrate；
Preferably, the substrate is transparent substrates, preferably any in PMMA substrate, PS substrate, COP substrate or COC substrate
It is a kind of or at least two combination.
8. the micro-fluidic chip described in one of -7 based on immune detection and biochemistry detection according to claim 1, which is characterized in that
Including substrate, sample injection unit, immune detection area and biochemistry detection area is arranged in substrate；
Sample injection unit includes sample introduction tank, the first quantitative slot, collecting tank and the first distributing trough, sample introduction tank, the first quantitative slot and collection
Slot passes sequentially through microchannel connection, and the first quantitative slot is also connected to the first distributing trough；First quantitative slot is connected to the first distributing trough
Microchannel on the first valve is set；
Immune detection area includes the second quantitative slot being sequentially communicated, the second allocation unit, at least one first reaction member and gives up
Liquid recovery unit；Further include the first liquid storage unit, solidification oxidant layer is provided in the first reaction member；
Second allocation unit includes the second distributing trough；First reaction member includes that third quantitative slot, the first mixing channel and first are anti-
Answer slot；First liquid storage unit includes fluid storage tanks；Devil liquor recovery unit includes waste liquid tank；
Second distributing trough, third quantitative slot, the first mixing channel, the first reactive tank and waste liquid tank, wherein the second quantitative slot and first
Distributing trough connection；Fluid storage tanks are connected to the second distributing trough and/or third quantitative slot；Second quantitative slot and the second distributing trough connect
Second valve is set on logical microchannel, the 4th valve is set on the microchannel that third quantitative slot is connected to the first mixing channel, first is anti-
Third valve is set on the microchannel for answering slot to be connected to waste liquid tank；
Biochemistry detection area includes that the 4th quantitative slot being sequentially communicated, the second mixed cell, third allocation unit and the second reaction are single
Member；It further include second liquid storage unit, second liquid storage unit is connected to the second mixed cell, is set in the second reaction member
It is equipped with the second solidification oxidant layer；
Second mixed cell includes the second mixing channel, and third allocation unit includes third distributing trough, and the second reaction member includes the
Two reactive tanks；Second liquid storage unit includes fluid storage tanks；
4th quantitative slot is connected to the first distributing trough, and the 5th is arranged on the microchannel that the 4th quantitative slot is connected to the second mixing channel
The 6th valve is arranged on the microchannel that the second mixing channel is connected to third distributing trough for valve.
9. a kind of detector based on immune detection and biochemistry detection, which is characterized in that the detector includes claim 1-8
One of described in micro-fluidic chip；
Preferably, the detector based on immune detection and biochemistry detection further includes sensing chamber, centrifugal device, light source, light letter
Number reception device, light signal processing device and display device；The substrate is fixed in centrifugal device, and is located in sensing chamber；
Light source is set in sensing chamber, is connected with display device；Light signal receiving is set in sensing chamber, passes through optical signal prosessing
Device is connected with display device；
Preferably, the detector based on immune detection and biochemistry detection further includes optical signal enhancement device, preferably photoelectricity
Multiplier tube, light signal receiving are connected by optical signal enhancement device with light signal processing device；
Preferably, the light signal receiving includes light-receiving probe.
10. a kind of method that immune detection and biochemistry detection are carried out using micro-fluidic chip described in one of claim 1-8,
It is characterized in that, described method includes following steps:
(1) sample to be tested is added, by rotating micro-fluidic chip sample to be tested is flowed；
(2) rotation micro-fluidic chip makes sample to be tested flow into immune detection area and biochemistry detection area, is immunoreacted respectively
And biochemical reaction；
(3) rotation micro-fluidic chip makes the liquid discharge after immune response.
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| CN109999931A (en) * | 2019-04-18 | 2019-07-12 | 天津诺迈科技有限公司 | Chemiluminescence detection micro-fluidic chip and application method, reagent cleaning method |
| CN110412256A (en) * | 2018-12-28 | 2019-11-05 | 苏州康容生物医疗科技有限公司 | A kind of blood sample test chip apparatus |
| CN110568202A (en) * | 2019-09-12 | 2019-12-13 | 重庆科技学院 | automatic sample separation constant volume immunofluorescence quantitative rapid detection micro-fluidic chip |
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| CN111855994A (en) * | 2020-07-29 | 2020-10-30 | 成都微康生物科技有限公司 | POCT (point of care testing) immunodetection chip capable of carrying out multiple joint detections on whole blood sample adding at one time |
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