CN107328931A - A kind of quick continuous detection technique based on nano-probe and magnetic micro-nano granules - Google Patents
A kind of quick continuous detection technique based on nano-probe and magnetic micro-nano granules Download PDFInfo
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- CN107328931A CN107328931A CN201710340689.6A CN201710340689A CN107328931A CN 107328931 A CN107328931 A CN 107328931A CN 201710340689 A CN201710340689 A CN 201710340689A CN 107328931 A CN107328931 A CN 107328931A
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Classifications
-
- 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
-
- 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
- G01N33/54346—Nanoparticles
Abstract
The present invention proposes a kind of while realizing multiple protein, nucleic acid or pathogen using nano-probe, paramagnetism micro-nano granules and micro-fluidic chip, quick and continuous detection.The magnetic micro-nano granules that the nano-probe and antibody or nucleic acid modified using monoclonal antibody or complementary nucleic acid sequences are modified are used as detection reagent, detection reaction tank is used as using multichannel micro-fluidic chip, the magnetic micro-nano granules that antibody or nucleic acid are modified are fixed on reaction tank bottom with magnetic field and form " erasable detection substrate ", using UV, visible light light splitting, fluorescence or Electrochemical detector as signal pickup assembly, while realizing multiple protein, nucleic acid or pathogen, quick detection.Have benefited from the erasable property of " nano immune substrate " and the anti-adhesive properties of chip material, the detector can realize the continuous detection of different samples, substantially increase the automation of detection and handle the ability of sample size, will be with a wide range of applications in fields such as medical diagnosis on disease, infectious disease monitoring and food sanitation safe monitorings.
Description
Technical field
The invention belongs to biochemical analysis technical field, and in particular to a kind of based on nano-probe and magnetic micro-nano granules
Quick continuous detection technique.
Background technology
The detection of protein, nucleic acid and pathogen is in necks such as medical diagnosis on disease, infectious disease monitoring and food sanitation safe monitorings
Domain is with a wide range of applications.Conventional detection method includes at present：（1）Test in laboratory based on large-scale instrument, it has
There is an advantage that accuracy is high, but it is detection cycle length, big to the consumption of manpower and need stored sample；（2）Based on the small of scene
Typeization is detected immediately（Point-of-care Testing, POCT）The detection of equipment, it is ensureing certain sensitivity and accuracy
While have the advantages that portability, instant quick detection and automation；（3）Detection based on test strips, it has user
Just, the low advantage of price, but accuracy and quantitation capabilities are not enough.To sum up, the detection technique based on scene miniaturization POCT equipment
There is great prospect in biochemical analysis field.
POCT be laboratory medicine development frontier, wherein home-use blood glucose meter be POCT be applied to chronic disease management into
Work(example；And constantly expanded using scope, can be applied to clinical examination, chronic disease detection, infectious disease detection, emergent anti-terrorism,
The public health such as food security, illicit drugs inspection, security fields.Detected in China POCT detection species comprising infectious disease and be
The eugenic detection series of row, gestation, lesion detection series, illicit drugs inspection series, digital blood glucose meter detection series, electronic sphygmomanometer system
Row etc., also clenbuterol hydrochloride quick detection series of products can directly detect in pork whether situation containing clenbuterol hydrochloride；And near
The development of several years automatic technologies and immunological technique, POCT is widely used in immune detection.In the market is most widely used
POCT technologies be immunochromatography and diafiltration techniques, in addition also include dry chemical technology, multilayer film technology, micro-fluidic skill
Art and be not required to blood sampling, can it is transdermal monitoring blood glucose and hemoglobin etc. infrared and far infrared spectrophotometry；In addition, selectivity
Fuzzy identification technology of electrode technology, biochip and biology sensor, microscope imaging etc., its test object also by it is immune,
Biochemistry progressively develops into detection of nucleic acids.And P/PC R detection reagents use also increasingly increase with equipment and have gone on market.
POCT from it is qualitative to sxemiquantitative again to accurate quantification be development inexorable trend, qualitative aspect immunochromatography technique is POCT
Successful model；At quantitative aspect, detection device and reagent have been realized in accurate quantification, with conventional fluorescent, time resolution skill
Based on a variety of luminescence technologies such as art, electrochemical luminescence and enzymology be luminous.But POCT apparatus fields, mesh are minimized at the scene
Before also lack can rapidly, continuously, high-throughout detection technique, it is contemplated that by proposing a kind of to be based on nano-probe and magnetic
The detection technique of property micro-nano granules, a kind of possible settling mode is provided for this major issue.
Micro-fluidic chip（Microfluidic chip）Analysis system is the new and high technology that develops rapidly at present and multidisciplinary
One of International Technology Disciplinary Frontiers of intersection, its essential characteristic and sharpest edges are a variety of monotechnicss overall controllable small
Scale is integrated on platform, flexible combination, can be integrated in the unit involved by biochemical analysis on very small chip,
To a kind of detection technique platform for the various functions for replacing conventional chemical or biology laboratory.Compared with conventional analytical techniques,
Micro-fluidic chip platform have flexible design, efficiently separate, quickly analysis, low reagent consumption, equipment miniaturization and it is highly integrated
The features such as change；And related generally to the almost all of analysis such as food hygiene, medical separation, environmental analysis, biochemical analysis
Detection field.
By the way that nano-probe, magnetic micro-nano granules and micro-fluidic chip are combined, the present invention proposes a kind of new
Continuous Fast Detection Technique.
The content of the invention
Be based on nano-probe it is an object of the invention to provide one kind, and combine magnetic micro-nano granules with it is micro-fluidic continuous
Fast Detection Technique.
To achieve the above object, the present invention provides a kind of continuous quick inspection based on nano-probe and magnetic micro-nano granules
Survey platform, including nano-probe, magnetic micro-nano granules detection substrate and micro-fluidic chip；
Described micro-fluidic chip includes microfluidic control passage and control system, and it is to be checked that described microfluidic control passage includes first
Sample entrance, second treat that sample entrance, the outlet of the first microfluidic channel, the outlet of the second microfluidic channel, the 3rd microfluidic channel go out
Mouth, the outlet of the 4th microfluidic channel, magnetic nanoparticle entrance, buffer inlet, the first detection zone, the second detection zone,
First microfluidic channel is exported to be connected with the outlet of the second microfluidic channel by the first microchannel, and is sequentially connected first
Sample entrance and the first detection zone are treated,
3rd microfluidic channel is exported to be connected with the outlet of the 4th microfluidic channel by the second microchannel, and is sequentially connected second
Sample entrance and the second detection zone are treated,
First microchannel is connected to be connected with the second microchannel and is connected by the 3rd microchannel, the 3rd described microchannel
Magnetic nanoparticle entrance and buffer inlet are connected simultaneously,
First treats sample entrance and the first microchannel connecting place provided with valve, and second treats that sample entrance and the second microchannel are connected
Place is provided with valve, and magnetic nanoparticle entrance and the 3rd microchannel connecting place are provided with valve, buffer inlet and the 3rd microchannel
Connecting place is provided with valve, and the first detection zone and the second detection zone are provided with magnetic field sources,
UV, visible light light splitting, fluorescence and Electrochemical detector can be used to be used as signal for the first described detection zone and the second detection zone
Harvester.
The magnetic micro-nano granules detection substrate is first to receive monoclonal antibody or complementary nucleic acid sequences modification in magnetic
Rice grain surface, then be passed through microfluidic channel, and by apply magnetic field so be fixed on detection zone being capable of specific recognition mesh
Mark the detection substrate of thing.In order to realize quick detection while multiple objects, it can prepare and be repaiied by different antibodies or nucleic acid fragment
The magnetic micro-nano granules adornd, and in the presence of microfluidic channel and magnetic field, in the detection zone bottom of material not of the same race point
Substrate Xing Cheng not be detected accordingly；
The nano-probe refers to by the fluorescent nano particle of monoclonal antibody or nucleic acid modified, with ultravioletvisible absorption
Nano particle and the nano particle with electro-chemical activity, and being capable of the corresponding object of specific recognition.It is multiple in order to realize
Detected while object, prepared nano-probe should have different fluorescence emission spectrums, different ultravioletvisible absorptions
Peak or different electrochemical redox current potentials.Simultaneously by preparing the nano particle by different antibodies or nucleic acid modified, shape
The specific recognition of paired different target thing.
The micro-fluidic chip uses the preparations such as Teflon or glass.
It is described at the same the object of detection can be it is a kind of, two or more.
The relevant designs such as the passage and detection zone of micro-fluidic chip can change for quantity, species of target detection thing etc..
The continuous quick detection side of a kind of nano-probe, magnetic micro-nano granules and micro-fluidic chip is further provided
Method, comprises the following steps：
First, erasable detection substrate is formed：
Step 1）Apply magnetic field in micro-fluidic chip detection zone bottom；
Step 2）The magnetic micro-nano granules that surface modification has antibody or nucleic acid are passed through in microfluidic channel；
Step 3）Buffer solution is passed through in microfluidic channel, micro-nano of the magnetic that detection zone bottom is not fixed on by magnetic field is rinsed
Grain, and then form erasable detection substrate；
Secondly, capture object and nano-probe：
Step 4）Nano-probe is mixed with sample to be tested, the object in sample is fully combined with nano-probe；
Step 5）By step 4）Mixed liquor be passed through microfluidic channel, the detection substrate of detection zone can be by the antibody of surface modification
Or nucleic acid capture object, nano-probe is now combined on object, i.e., forms sandwich knot on the detection zone surface of passage
Structure；
Step 6）Buffer solution is passed through in microfluidic channel, the material for not being detected substrate capture is rinsed；
Then, detected：
Step 7）The detection method that should be used according to micro-fluidic chip detection zone, can detect ultraviolet-visible absorption, fluorescence intensity
Or electrochemical signals, carry out data processing；
Finally, clean chip channel and initially enter the detection of next sample：
Step 8）Close magnetic field；
Step 9）Buffer solution is passed through in microfluidic channel, all substances in microfluidic channel are rinsed（Mainly include magnetic micro-nano
Rice grain detection substrate, object and nano-probe）；
Step 10）Carry out next sample detection.
Used sample can be blood, urine, saliva, food extract solution equal samples or dilution or other may contain
There is the sample of target detection thing.
Step 4）Incubation time in middle nano-probe and sample required for the combination of object by object and probe it
Between affinity and the factor such as properties of samples determine.
The present invention provide based on nano-probe, magnetic micro-nano granules and micro-fluidic continuous Fast Detection Technique, tool
There is following remarkable advantage：
1st, the present invention by using with different fluorescence emission spectrums, different ultraviolet and visible absorption peak and different electrochemical oxidations also
The nano-probe of former current potential, is detected while can realizing plurality of target thing, improves the flux of detection；
2nd, the magnetic micro-nano granules " detection substrate " modified by antibody or nucleic acid in the present invention are by magnetic field control, with erasable
The property write, makes the detector to realize the continuous detection of different samples, substantially increases the automation of detection and handles sample size
Ability；
3rd, micro-fluidic chip has anti-adhesive properties in the present invention, helps realize the continuous detection of different samples；
4th, the present invention incorporates specific marker and nano material carries out joint-detection, and is combined with microchannel, Neng Goushi
Quick detection while existing plurality of target thing, simplifies testing process, the cost-effective and time.
Brief description of the drawings
Fig. 1 shows detection technique principle schematic of the present invention.
Fig. 2 is shown detects two kinds of marker of inflammation PCT and CRP while a kind of exemplary embodiment of the invention is proposed
Microfluidic channel schematic diagram.
Wherein, 1 is that valve 1,2 is that valve 2,3 is that valve 3,4 is magnetic field sources, 5 is first to treat that sample entrance, 6 are second to treat that sample enters
Mouthful, 7 be magnetic nanoparticle entrance, 8 be buffer inlet, 9 be microfluidic channel outlet, 10 be the first detection zone, 11 be the
Two detection zones.
Embodiment
Embodiment 1：A kind of albumen based on rare-earth fluorescent nano-probe, magnetic micro-nano granules and micro-fluidic chip is continuous
Quick detection platform
Microfluidic channel schematic diagram involved by the present embodiment is as shown in Figure 2.In the following description, " magnetic nanoparticle 1 "
The magnetic nanoparticle of the monoclonal antibody of surface modification albumen 1 is represented, " magnetic nanoparticle 2 " represents surface modification albumen
The magnetic nanoparticle of 2 monoclonal antibodies, " rare earth nano probe 1 " represents the rare earth of the monoclonal antibody of surface modification albumen 1
Nano-probe, " rare earth nano probe 2 " represents the rare earth nano probe of the monoclonal antibody of surface modification albumen 2.Its middle rare earth
Nano-probe can be the rare-earth fluorescent nano particle based on elements such as such as Eu, Gd, Tb, Dy and Yb.
First, nano immune substrate is formed：
Step 1）Apply magnetic field in micro-fluidic chip detection zone bottom；
Step 2）The magnetic nanoparticle 1,2 of certain volume is passed through in microfluidic channel；
Step 3）Buffer solution is passed through in microfluidic channel, the magnetic Nano for not being fixed on detection zone bottom by magnetic field is rinsed
Grain, and then form nano immune substrate；
Secondly, capture albumen 1, albumen 2 and rare earth nano probe 1,2：
Step 4）The rare earth nano probe 1 of certain volume is mixed with appropriate sample to be tested, make albumen 1 in sample fully with
Rare earth nano probe 1 is combined；Rare earth nano probe 2 is mixed with sample to be tested, makes albumen 2 in sample fully and rare earth nano
Probe 2 is combined；
Step 5）By step 4）Mixed liquor be passed through microfluidic channel treat sample entrance 1,2, the immune substrate of the detection zone of albumen 1
Can be by the antibody capture albumen 1 of surface modification, the immune substrate of the detection zone of albumen 2 can be by the antibody capture egg of surface modification
White 2;Now albumen 1 on albumen 2 on the detection zone surface of passage with respectively in connection with rare-earth fluorescent nano-probe 1,2, i.e., distinguishing
Form sandwich structure；
Step 6）Buffer solution is passed through in microfluidic channel, the detection zone of albumen 1 is rinsed with the detection zone of albumen 2 not by nano immune base
The material of bottom capture；
Then, detected：
Step 7）The excitation wavelength that micro-fluidic chip albumen 1 and the detection zone of albumen 2 should be used is determined, and detects that it launches light intensity
Degree, carries out data processing；
Finally, clean chip and initially enter the detection of next sample：
Step 8）Close magnetic field；
Step 9）Buffer solution is passed through in microfluidic channel, all substances in microfluidic channel are rinsed（Mainly include magnetic Nano
Particle 1,2, rare earth nano probe 1,2 and albumen 1,2 etc.）；
Step 10）Carry out next pattern detection.
Embodiment 2：A kind of nucleic acid based on gold nanoparticle probes, magnetic micro-nano granules and micro-fluidic chip is continuously fast
Fast detection platform
In the following description, " magnetic nanoparticle 1 " represents the magnetic Nano of the complementary series of surface modification nucleic acid 1 to the present embodiment
Particle, " magnetic nanoparticle 2 " represents the magnetic nanoparticle of the complementary series of surface modification nucleic acid 2, " the generation of gold nano-probe 1 "
The table surface modification gold nanoparticle probes of the complementary series of nucleic acid 1, " it is mutual that gold nano-probe 2 " represents surface modification nucleic acid 2
The gold nanoparticle probes of complementary series.Wherein, gold nano-probe can have difference with different ultraviolet and visible absorption peaks
Shape（Such as gold nano grain, gold nano disk and gold nano triangle）Or various sizes of gold nano grain.
First, nano nucleic acid detection substrate is formed：
Step 1）Apply magnetic field in micro-fluidic chip detection zone bottom；
Step 2）The magnetic nanoparticle 1,2 of certain volume is passed through in microfluidic channel；
Step 3）Buffer solution is passed through in microfluidic channel, the magnetic Nano for not being fixed on detection zone bottom by magnetic field is rinsed
Grain, and then form nano nucleic acid detection substrate；
Secondly, capture nucleic acid 1, nucleic acid 2 and gold nano-probe 1,2：
Step 4）The gold nano-probe 1 of certain volume is mixed with appropriate sample to be tested, make nucleic acid 1 in sample fully with gold
Nano-probe 1 is combined；Gold nano-probe 2 is mixed with sample to be tested, make nucleic acid 2 in sample fully with the knot of gold nano-probe 2
Close；
Step 5）By step 4）Mixed liquor be passed through microfluidic channel treat sample entrance, the detection of nucleic acids base of the detection zone of nucleic acid 1
Bottom can catch nucleic acid 1 by the complementary series of nucleic acid 1 of surface modification, and the detection of nucleic acids substrate of the detection zone of nucleic acid 2 can pass through surface
The complementary series of nucleic acid 2 of modification catches nucleic acid 2;Now respectively in connection with gold nano-probe 1,2 on nucleic acid 1 and nucleic acid 2, i.e., in passage
Detection zone surface form sandwich structure respectively；
Step 6）Buffer solution is passed through in microfluidic channel, the detection zone of nucleic acid 1 is rinsed with the detection zone of nucleic acid 2 not by detection of nucleic acids base
The material of bottom capture；
Then, detected：
Step 7）The ultravioletvisible absorption intensity of micro-fluidic chip nucleic acid 1 and the detection zone of nucleic acid 2 is detected, data processing is carried out；
Finally, clean chip and initially enter the detection of next sample：
Step 8）Close magnetic field；
Step 9）Buffer solution is passed through in microfluidic channel, all substances in microfluidic channel are rinsed（Mainly include magnetic Nano
Particle 1,2, gold nano-probe 1,2 and nucleic acid 1,2 etc.）；
Step 10）Carry out next pattern detection.
Embodiment 3：It is a kind of based on heavy metal electro-chemical activity nano-probe, magnetic micro-nano granules and micro-fluidic chip
The continuous quick detection platform of nucleic acid
In the following description, " magnetic nanoparticle 1 " represents the magnetic Nano of the complementary series of surface modification nucleic acid 1 to the present embodiment
Particle, " magnetic nanoparticle 2 " represents the magnetic nanoparticle of the complementary series of surface modification nucleic acid 2, " heavy metal nano-probe
1 " represents the heavy metal nano-probe of the complementary series of surface modification nucleic acid 1, and " heavy metal nano-probe 2 " represents surface modification
The heavy metal nano-probe of the complementary series of nucleic acid 2.Wherein, heavy metal nano-probe can be containing the weight such as such as Cr, Fe, Ni and Cu
The nano particle of metallic element.Also, the upper and lower surface of micro-fluidic chip detection zone is evenly equipped with electrode in the present embodiment（Such as lead
Electric glass ITO and FTO etc.）.
First, nano nucleic acid detection substrate is formed：
Step 1）Apply magnetic field in micro-fluidic chip detection zone bottom；
Step 2）The magnetic nanoparticle 1,2 of certain volume is passed through in microfluidic channel；
Step 3）Buffer solution is passed through in microfluidic channel, the magnetic Nano for not being fixed on detection zone bottom by magnetic field is rinsed
Grain, and then form nano nucleic acid detection substrate；
Secondly, capture nucleic acid 1, nucleic acid 2 and heavy metal nano-probe 1,2：
Step 4）The heavy metal nano-probe 1 of certain volume is mixed with appropriate sample to be tested, makes the nucleic acid 1 in sample abundant
Combined with heavy metal nano-probe 1；Heavy metal nano-probe 2 is mixed with sample to be tested, make nucleic acid 2 in sample fully with again
Metallic nano detecting probe 2 is combined；
Step 5）By step 4）Mixed liquor be passed through microfluidic channel treat sample entrance, the detection of nucleic acids base of the detection zone of nucleic acid 1
Bottom can catch nucleic acid 1 by the complementary series of nucleic acid 1 of surface modification, and the detection of nucleic acids substrate of the detection zone of nucleic acid 2 can pass through surface
The complementary series of nucleic acid 2 of modification catches nucleic acid 2;Now respectively in connection with gold nano-probe 1,2 on nucleic acid 1 and nucleic acid 2, i.e., in passage
Detection zone surface form sandwich structure respectively；
Step 6）Buffer solution is passed through in microfluidic channel, the detection zone of nucleic acid 1 is rinsed with the detection zone of nucleic acid 2 not by detection of nucleic acids base
The material of bottom capture；
Then, detected：
Step 7）The current strength of micro-fluidic chip nucleic acid 1 and the detection zone of nucleic acid 2 is detected, data processing is carried out；
Finally, clean chip and initially enter the detection of next sample：
Step 8）Close magnetic field；
Step 9）Buffer solution is passed through in microfluidic channel, all substances in microfluidic channel are rinsed（Mainly include magnetic Nano
Particle 1,2, heavy metal nano-probe 1,2 and nucleic acid 1,2 etc.）；
Step 10）Carry out next pattern detection.
Described above is only the preferred embodiment of the present invention, is not intended to limit the invention, for the technology of this area
For personnel, under the premise without departing from the principles of the invention, any modification, equivalent substitution and improvements made and retouching etc.,
It should be included within protection scope of the present invention.
Claims (10)
1. a kind of continuous quick detection platform based on nano-probe and magnetic micro-nano granules, it is characterised in that：Including nanometer
Probe, magnetic micro-nano granules detection substrate and micro-fluidic chip；
Described micro-fluidic chip includes microfluidic control passage and control system（As shown in Figure 2）, described microfluidic control passage
Treat that sample entrance, second treat sample entrance, the outlet of the first microfluidic channel, the outlet of the second microfluidic channel, the 3rd including first
Microfluidic channel outlet, the outlet of the 4th microfluidic channel, magnetic nanoparticle entrance, buffer inlet, the first detection zone, second
Detection zone,
First microfluidic channel is exported to be connected with the outlet of the second microfluidic channel by the first microchannel, and is sequentially connected first
Sample entrance and the first detection zone are treated,
3rd microfluidic channel is exported to be connected with the outlet of the 4th microfluidic channel by the second microchannel, and is sequentially connected second
Sample entrance and the second detection zone are treated,
First microchannel is connected to be connected with the second microchannel and is connected by the 3rd microchannel, the 3rd described microchannel
Magnetic nanoparticle entrance and buffer inlet are connected simultaneously,
First treats sample entrance and the first microchannel connecting place provided with valve, and second treats that sample entrance and the second microchannel are connected
Place is provided with valve, and magnetic nanoparticle entrance and the 3rd microchannel connecting place are provided with valve, buffer inlet and the 3rd microchannel
Connecting place is provided with valve, and the first detection zone and the second detection zone are provided with magnetic field sources.
2. the continuous quick detection platform according to claim 1 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：UV, visible light light splitting, fluorescence and Electrochemical detector conduct can be used in the first described detection zone and the second detection zone
Signal pickup assembly.
3. the continuous quick detection platform according to claim 1 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：The magnetic micro-nano granules detection substrate is first by monoclonal antibody or complementary nucleic acid sequences modification in magnetic Nano
Particle surface, then be passed through microfluidic channel, and by apply magnetic field so be fixed on detection zone being capable of specific recognition target
The detection substrate of thing；
In order to realize quick detection while multiple objects, it can prepare micro- by the magnetic of different antibodies or nucleic acid fragment modified
Nano particle, and in the presence of microfluidic channel and magnetic field, formed respectively accordingly in the detection zone bottom of material not of the same race
Detect substrate.
4. the continuous quick detection platform according to claim 1 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：The nano-probe refers to inhale by the fluorescent nano particle of monoclonal antibody or nucleic acid modified, with UV, visible light
The nano particle of receipts and the nano particle with electro-chemical activity, and being capable of the corresponding object of specific recognition；In order to realize
Detected while multiple objects, prepared nano-probe should have different fluorescence emission spectrums, different UV, visible lights
Absworption peak or different electrochemical redox current potentials, while by preparing the nanometer by different antibodies or nucleic acid modified
Grain, forms the specific recognition to different target thing.
5. the continuous quick detection platform according to claim 1 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：It is characterized in that：The micro-fluidic chip uses the preparations such as Teflon or glass.
6. the continuous quick detection platform according to claim 1 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：It is described at the same the object of detection can be it is a kind of, two or more.
7. the continuous quick detection platform according to claim 1 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：The relevant designs such as the passage and detection zone of micro-fluidic chip can change for quantity, species of target detection thing etc..
8. the continuous quick determination method of a kind of nano-probe, magnetic micro-nano granules and micro-fluidic chip, it is characterised in that：Bag
Include following steps：
First, erasable detection substrate is formed：
Step 1）Apply magnetic field in micro-fluidic chip detection zone bottom；
Step 2）The magnetic micro-nano granules that surface modification has antibody or nucleic acid are passed through in microfluidic channel；
Step 3）Buffer solution is passed through in microfluidic channel, micro-nano of the magnetic that detection zone bottom is not fixed on by magnetic field is rinsed
Grain, and then form erasable detection substrate；
Secondly, capture object and nano-probe：
Step 4）Nano-probe is mixed with sample to be tested, the object in sample is fully combined with nano-probe；
Step 5）By step 4）Mixed liquor be passed through microfluidic channel, the detection substrate of detection zone can be by the antibody of surface modification
Or nucleic acid capture object, nano-probe is now combined on object, i.e., forms sandwich knot on the detection zone surface of passage
Structure；
Step 6）Buffer solution is passed through in microfluidic channel, the material for not being detected substrate capture is rinsed；
Then, detected：
Step 7）The detection method that should be used according to micro-fluidic chip detection zone, can detect ultraviolet-visible absorption, fluorescence intensity
Or electrochemical signals, carry out data processing；
Finally, clean chip channel and initially enter the detection of next sample：
Step 8）Close magnetic field；
Step 9）Buffer solution is passed through in microfluidic channel, all substances in microfluidic channel are rinsed（Mainly include magnetic micro-nano
Rice grain detection substrate, object and nano-probe）；
Step 10）Carry out next sample detection.
9. the continuous quick determination method according to claim 8 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：Used sample can be blood, urine, saliva, food extract solution equal samples or dilution or other may contain
There is the sample of target detection thing.
10. the continuous quick determination method according to claim 8 based on nano-probe and magnetic micro-nano granules, it is special
Levy and be：Step 4）Incubation time in middle nano-probe and sample required for the combination of object is between object and probe
Affinity and the factor such as properties of samples determine.
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