CN110261602A - A kind of detection method and detection kit based on fluorescence-encoded magnetic bead - Google Patents
A kind of detection method and detection kit based on fluorescence-encoded magnetic bead Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
- G01N15/10—Investigating individual particles
- G01N15/14—Optical investigation techniques, e.g. flow cytometry
- G01N15/1434—Optical arrangements
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- G—PHYSICS
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- 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
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
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- 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
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- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
- G01N33/582—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances with fluorescent label
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- 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
- G01N33/58—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving labelled substances
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Abstract
The invention belongs to magnetic bionanoparticles application and technical field of medical examination, in particular to a kind of detection method based on fluorescence-encoded magnetic bead, including capturing step, form compound step, separating step and excitation determination step, the capture step, compound step and separating step are formed using target molecule needed for fluorescence-encoded micro-beads specific binding capture, means of transport using magnetic posts as compound, it shifts in different reaction environments, automation can be used to be controlled, realize quickly detection, fluorescence spectrophotometer detection can be used in the excitation and determination step, FCM analysis method and liquid-phase chip detection method, detection and high-throughput detection while realization to different target molecule, especially FCM analysis method, it can be by storing fluorescence-encoded micro-beads spectral waveform database, realize intelligent automation detection.
Description
Technical field
The invention belongs to magnetic bionanoparticles application and technical field of medical examination, in particular to a kind of based on fluorescence-encoded
The detection method and detection kit of magnetic bead.
Background technique
The object of clinical medical inspection includes mainly cell class and small-molecule substance two major classes, and small-molecule substance includes egg
White matter or nucleic acid etc..The detecting instrument of cell sample common are flow cytometer, cellanalyzer etc., and small-molecule substance
Detecting instrument then include chemiluminescence detector, chromatograph etc..In actual application, the detection of small-molecule substance,
Such as tumor markers, common detection mode be chemoluminescence method carry out individual event detection, but from clinical meaning for, tumour
Marker and tumour are not the corresponding relationships in complete meaning, only a kind of correlation, the detection of individual event tumor markers, tumour
Recall rate is low, and multi objective parallel detection and diagnosis, increases the sensibility and specificity of lesion detection, substantially increases tumour
Recall rate, similarly, the joint-detection of other a variety of small-molecule substances is also of great significance for the diagnosis of clinical disease.
In recent years, someone combines chemiluminescence with nanotechnology, proposes chemiluminescence magnetic enzyme immunoassay (EIA), will
Magnetic separation technique, immunological method are combined with chemiluminescence detection technology three, and the complexity of antibody cladding is greatly saved
Process simplifies operation, saves the time.It is mixed to have developed center according to the producing principle of micro-fluidic chip by scholar Lin Jinming etc.
The three flow path chemiluminescence flow injection chip detection cells closed, realize the in situ detection of three kinds of substances, however, facing more multiple groups
Point, or the joint-detection of more micro substance, then it is difficult to realize accurate sensitive detection.
With the development of full-automatic, big data and artificial intelligence technology, simplify detection process, realizes that multicomponent automation is same
When detection at a kind of development trend, and the prior art is difficult to realize to high-throughput various ingredients automation simultaneously as described above
Detection.
Summary of the invention
Therefore, the technical problem to be solved in the present invention is that the prior art can not achieve high throughput, multicomponent, automation inspection
It surveys, detection method is complicated, and more disadvantage at high cost provides a kind of detection method based on fluorescence-encoded magnetic bead.
The invention discloses a kind of detection methods based on fluorescence-encoded magnetic bead, include the following steps
Capture step: fluorescence-encoded magnetic bead is mixed with the product to be tested containing target molecule, captures target molecule；
It forms compound step: fluorescent reporter group is added, specificity is connected on captured target molecule, forms fluorescence
Encoded magnetic bead-target molecule-fluorescent reporter group compound；
Separating step: adsorbing the fluorescence-encoded magnetic bead-target molecule-fluorescent reporter group compound using magnetic posts,
And it is transferred in subsequent reactions environment；
Excitation and determination step: it is multiple that the fluorescence-encoded magnetic bead-target molecule-fluorescent reporter group is irradiated using exciting light
Object is closed, the fluorescence-encoded magnetic bead-target molecule-fluorescent reporter group compound is identified according to spectral waveform, according to
The fluorescence intensity of fluorescent reporter group carries out quantitative detection to captured target molecule.
Preferably, the magnetic posts are column structure, and magnetic posts working end diameter is 1mm-5cm, by current control
Magnetic state.
Preferably, the reaction environment includes buffer, confining liquid or sheath fluid.
Preferably, further include sample pre-treatments step, target molecule in sample is separated and is enriched with before capturing step.
It preferably, further include the preparation of fluorescence-encoded magnetic bead mixed liquor before capturing step, using at least two different fluorescence
The fluorescence-encoded magnetic bead mixing of coding capture while at least two target molecules.
Preferably, it is described capture step further include capture target molecule after elution and closing step and formed compound
Elution and closing step after forming the fluorescence-encoded magnetic bead-target molecule-fluorescent reporter group compound in step.
Preferably, the elution and closing step are adsorbed and are shifted using magnetic posts.
Preferably, the detecting instrument that the excitation and determination step use is selected from sepectrophotofluorometer, flow cytometer
Or liquid phase suspension array chip.
The invention also discloses a kind of detection kits using the detection method, including,
It is coated with the fluorescence-encoded micro-beads of target molecule；
The detection antibody of fluorescent reporter group label；
Negative controls；
Positive reference substance；
Buffer.
Preferably, fluorescence signal molecule and the fluorescent reporter group are selected from center transmitting on the fluorescence-encoded micro-beads
The different CdSe/ZnS quantum dot of wavelength, the CdSe/ZnS quantum dot center emission wavelength are 540-680nm.
The invention also discloses based on fluorescence-encoded magnetic bead described in one kind detection method or the detection kit glimmering
The application of light analysis field.
Technical solution of the present invention has the advantages that
1. the detection method of the present invention based on fluorescence-encoded magnetic bead, including capture step, form compound step, divide
From step and excitation determination step, the capture step forms compound step and separating step using fluorescence-encoded micro-beads spy
Target molecule needed for the opposite sex combines capture, the means of transport using magnetic posts as compound are shifted in different reaction rings
Border can be used automation and be controlled, and realize quickly detection, and fluorescence spectrophotometer detection, stream can be used in the excitation and determination step
Formula cell assay and liquid-phase chip detection method, detection and high-throughput detection while realization to different target molecule, especially
FCM analysis method can realize intelligent automation detection by storing fluorescence-encoded micro-beads spectral waveform database.
2. the detection method of the present invention based on fluorescence-encoded magnetic bead is passed through using magnetic posts as transfer medium
Electric current controls the presence or absence of magnetic posts magnetism and magnetic size, magnetic absorption action temperature and, biomolecule is influenced it is small, and
Movement by controlling magnetic posts can be achieved with the transfer of fluorescence-encoded micro-beads, is not required to artificial treatment, is advantageously implemented automation
And detection efficiency.
3. the detection method of the present invention based on fluorescence-encoded magnetic bead, including after capture target molecule elution and
Close step and form the elution and closing step in compound step, can using magnetic posts as transfer medium, with
Realize the automation of entire detection process.
4. the detection that the detection method of the present invention based on fluorescence-encoded magnetic bead, the excitation and determination step use
Instrument is selected from sepectrophotofluorometer, flow cytometer or liquid phase suspension array chip, in conjunction with magnetic posts, realizes automation, height
Flux and quickly detection.
Specific embodiment
There is provided following embodiments is to preferably further understand the present invention, it is not limited to the best embodiment party
Formula is not construed as limiting the contents of the present invention and protection scope, anyone under the inspiration of the present invention or by the present invention and its
The feature of his prior art is combined and any and identical or similar product of the present invention for obtaining, all falls within of the invention
Within protection scope.
Specific experiment step or condition person are not specified in embodiment, according to the literature in the art described routine experiment
The operation of step or condition can carry out.Reagents or instruments used without specified manufacturer, being can be by commercially available acquisition
Conventional products.
Embodiment 1
A kind of detection kit is present embodiments provided, including 1) is coated with the anti-human alpha-fetoprotein of mouse (AFP) monoclonal antibody
Fluorescence-encoded micro-beads；2) quantum dot-labeled detection antibody；3) positive reference substance；4) negative controls；5) pH7.2-7.4
Phosphate buffer.
Wherein, the fluorescence-encoded micro-beads are 2%Fe to be embedded with mass fraction3O4Superparamagnetic nanoparticle, partial size are
The polystyrene microsphere of 15-20um, surface are coated with 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (EDC)
With N- hydroxy thiosuccinimide (NHS), can be formed in conjunction with the amino in the anti-human alpha-fetoprotein monoclonal antibody of mouse altogether
Valence link, having on the fluorescence-encoded micro-beads be using amino modified center hair for fluorescence-encoded fluorescence signal molecule
The CdSe/ZnS quantum dot of ejected wave a length of 560nm and 630nm, the mass ratio of the CdSe/ZnS quantum dot of two kinds of center emission wavelengths
For 1:1.
The quantum dot-labeled detection antibody is the anti-human alpha-fetoprotein of mouse (AFP) monoclonal antibody, thereon saturation flags
Promising center emission wavelength is the CdSe/ZnS quantum dot of 680nm.
The positive reference substance is alpha-fetoprotein (AFP) standard items.
The negative controls are the blood serum sample without containing alpha-fetoprotein.
The present embodiment additionally provides a kind of detection method based on fluorescence-encoded magnetic bead, specifically includes the following steps:
Sample pre-treatments step: whole blood sample to be measured being carried out to be centrifuged 15min with 8000rpm, obtains serum as detection
Sample；Continuous ten times of standard items of the positive reference substance alpha-fetoprotein (AFP) are diluted, for use；
Capture step: taking 300ul fluorescence-encoded micro-beads reagent in 96 orifice plates, and parallel 3 parts, every part of 5 holes, first part of addition
Serum 10ul after centrifugation, second part of positive reference substance for being separately added into the diluted various concentration of 10ul, third part add respectively
Enter 10ul negative controls, 37 DEG C of shaking tables react 30min, and using diameter is that 5mm magnetic posts are fixed on fluorescence-encoded magnetic bead
It in magnetic posts, is then transferred in buffer with magnetic posts, controls electric current, make magnetic disappearance, the fluorescence for capturing target molecule is compiled
Code microballoon is scattered in buffer again, magnetic posts is adsorbed in after cleaning, then the PBS of the BSA containing 1%wt is transferred to magnetic posts
Same as above in confining liquid, dispersion is adsorbed in magnetic posts after closing；
It forms compound step: the magnetic posts for being adsorbed with fluorescence-encoded micro-beads being transferred to and are marked with quantum equipped with 500ul
In the detection antibody reagent of point, dispersion, 37 DEG C of reaction shaking tables form fluorescence-encoded magnetic bead-target molecule-fluorescence after reacting 30min
Reporter group compound, is adsorbed in magnetic posts；
Separating step: will be adsorbed with fluorescence-encoded micro-beads-target molecule-detection antibody complex magnetic posts be transferred to it is slow
Unreacted detection antibody and background material are eluted in fliud flushing, are then dispersed in buffer；
Excitation and determination step: using sepectrophotofluorometer, and wavelength is used to excite for the excitation wavelength of 618nm, detection
Fluorescence-encoded micro-beads-target molecule-detection antibody complex fluorescence in positive reference substance, negative controls and the buffer
Intensity is strong according to the quantum dot fluorescence marked on the fluorescence-encoded micro-beads-target molecule-detection antibody complex detection antibody
The quantum dot fluorescence strength criterion curve of degree and positive reference substance calculates the content of testing molecule in sample.
Embodiment 2
Detection kit described in the present embodiment and detection method are substantially the same manner as Example 1, and difference is only that, using stream
Fluorescence-encoded micro-beads-target molecule-in the formula cell instrument detection positive reference substance, the negative controls and the buffer
The fluorescence intensity of antibody complex is detected, the content of testing molecule in sample is calculated.
Embodiment 3
Present embodiments provide a kind of detection kit, including, 1) coating 5 kinds of different target molecules antibody fluorescence compile
Code microballoon；2) quantum dot-labeled 5 kinds corresponding detection antibody；3) 5 kinds of positive reference substances；4) negative controls；5)
The phosphate buffer of pH7.2-7.4；
The fluorescence-encoded micro-beads are 3%Fe to be embedded with mass fraction3O4Superparamagnetic nanoparticle, partial size 15-20um
Polystyrene microsphere, surface is coated with 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (EDC) and N- hydroxyl
Thiosuccimide (NHS) can form covalent bond, the packet in conjunction with the amino in the monoclonal antibody of target molecule
It is as follows by the fluorescence-encoded micro-beads information of the antibody of 5 kinds of different target molecules:
Fluorescence-encoded micro-beads are 1.: described for fluorescence-encoded fluorescence signal group be using amino modified center transmitting
Wavelength is the CdSe/ZnS quantum dot of 540nm, 570nm, 600nm and 630nm, the CdSe/ZnS quantum of 4 kinds of center emission wavelengths
The mass ratio of point is 1:1:1:1；Target molecule antibody on the fluorescence-encoded micro-beads is the anti-human alpha-fetoprotein of mouse (AFP) Dan Ke
Grand antibody, the quantum dot-labeled detection antibody are the anti-human alpha-fetoprotein of mouse (AFP) monoclonal antibody, the quantum marked thereon
Point is the CdSe/ZnS quantum dot that center launch wavelength is 680nm；
Fluorescence-encoded micro-beads are 2.: described for fluorescence-encoded fluorescence signal molecule be using amino modified center transmitting
Wavelength is the CdSe/ZnS quantum dot of 540nm, 570nm, 600nm and 630nm, the CdSe/ZnS quantum of 4 kinds of center emission wavelengths
The mass ratio of point is 1:2:1:3；Target molecule antibody on the fluorescence-encoded micro-beads is mouse anti-human prostate specific antigen
(PSA) monoclonal antibody, the quantum dot-labeled detection antibody are anti-for mouse anti-human prostate specific antigen (PSA) monoclonal
Body, the quantum dot marked thereon are the CdSe/ZnS quantum dot that center launch wavelength is 680nm；
Fluorescence-encoded micro-beads are 3.: described for fluorescence-encoded fluorescence signal molecule be using amino modified center transmitting
Wavelength is the CdSe/ZnS quantum dot of 540nm, 570nm, 600nm and 630nm, the CdSe/ZnS quantum of 4 kinds of center emission wavelengths
The mass ratio of point is 1:2:3:4；Target molecule antibody on the fluorescence-encoded micro-beads is the anti-human carcinomebryonic antigen of mouse (CEA) Dan Ke
Grand antibody, the quantum dot-labeled detection antibody are the anti-human carcinomebryonic antigen of mouse (CEA) monoclonal antibody, the quantum marked thereon
Point is the CdSe/ZnS quantum dot that center launch wavelength is 680nm；
Fluorescence-encoded micro-beads are 4.: described for fluorescence-encoded fluorescence signal molecule be using amino modified center transmitting
Wavelength is the CdSe/ZnS quantum dot of 540nm, 570nm, 600nm and 630nm, the CdSe/ZnS quantum of 4 kinds of center emission wavelengths
The mass ratio of point is 5:4:3:2；Target molecule antibody on the fluorescence-encoded micro-beads is the specific enol of mouse anti human nerve member
Change enzyme (NSE) monoclonal antibody, the quantum dot-labeled detection antibody is mouse anti human nerve member specificity olefinic alcohol enzyme (NSE)
Monoclonal antibody, the quantum dot marked thereon are the CdSe/ZnS quantum dot that center launch wavelength is 680nm；
Fluorescence-encoded micro-beads are 5.: described for fluorescence-encoded fluorescence signal molecule be using amino modified center transmitting
Wavelength is the CdSe/ZnS quantum dot of 540nm, 570nm, 600nm and 630nm, the CdSe/ZnS quantum of 4 kinds of center emission wavelengths
The mass ratio of point is 5:3:1:2；Target molecule antibody on the fluorescence-encoded micro-beads is the anti-human CA199 carbohydrate antigen Dan Ke of mouse
Grand antibody, the quantum dot-labeled detection antibody are the anti-human CA199 carbohydrate antigen monoclonal antibody of mouse, the quantum marked thereon
Point is the CdSe/ZnS quantum dot that center launch wavelength is 680nm；
The positive reference substance is alpha-fetoprotein (AFP) standard items, prostate specific antigen (PSA) standard items, cancer embryo are anti-
Former (CEA) standard items, through first specificity olefinic alcohol enzyme (NSE) standard items, CA199 carbohydrate antigen standard items.
The negative controls are the blood serum sample without containing target molecule.
The present embodiment additionally provides a kind of detection method based on fluorescence-encoded magnetic bead, specifically includes the following steps:
Sample pre-treatments step: whole blood sample to be measured being carried out to be centrifuged 15min with 8000rpm, obtains serum as detection
Sample；5 kinds of positive reference substance standard items are taken to mix, respectively continuous ten times of dilutions, for use；
Capture step: take respectively 100ul fluorescence-encoded micro-beads 1., 2., 3., 4., 5. reagent be mixed to form mixing magnetic bead in
In 96 orifice plates, parallel 3 parts, first part of serum 5ul being added after centrifugation, second part is separately added into the diluted various concentration of 5ul
Positive reference substance mixed liquor, third part is separately added into 5ul negative controls, and 37 DEG C of shaking tables react 30min, are using diameter
5cm magnetic posts are fixed on fluorescence-encoded magnetic bead in magnetic posts, are then transferred in buffer with magnetic posts, control electric current,
Making magnetic disappearance, the fluorescence-encoded micro-beads for capturing target molecule are scattered in buffer again, magnetic posts are adsorbed in after cleaning, then
It is transferred in the PBS confining liquid of the BSA containing 1%wt with magnetic posts, same as above, dispersion is adsorbed in magnetic posts after closing；
It forms compound step: the magnetic posts for being adsorbed with fluorescence-encoded micro-beads being transferred to and are marked with quantum equipped with 500ul
In the detection antibody reagent of point, dispersion, 37 DEG C of reaction shaking tables form fluorescence-encoded magnetic bead-target molecule-fluorescence after reacting 30min
Reporter group compound, is adsorbed in magnetic posts；
Separating step: will be adsorbed with fluorescence-encoded micro-beads-target molecule-detection antibody complex magnetic posts be transferred to it is slow
Unreacted detection antibody and background material are eluted in fliud flushing, are then dispersed in buffer；
Excitation and determination step: using flow cytometer, and excitation wavelength is the excitation of 488nm and 618nm, detection
Fluorescence-encoded micro-beads-target molecule-detection antibody complex fluorescence spectrum waveform and center emission wavelength in the buffer
For the fluorescence intensity of 680nm quantum dot, according to the reference spectra waveform in flow cytometer reference spectra waveform library to described glimmering
Pumped FIR laser microballoon-target molecule-detection antibody complex is classified, according to the quantum dot fluorescence intensity marked on detection antibody
The content of testing molecule in sample is calculated with the quantum dot fluorescence strength criterion curve of positive reference substance.
Embodiment 4
Detection kit described in the present embodiment and detection method are substantially the same manner as Example 3, and difference is only that, using liquid
Phase chip detects fluorescence-encoded micro-beads-target molecule-detection antibody complex fluorescence intensity, excitation wavelength in the buffer
For 488nm and 618nm, the fluorescence-encoded micro-beads-target molecule-detection antibody complex is divided according to spectral waveform
Class, according to the quantum dot fluorescence strength criterion curve of quantum dot fluorescence intensity and positive reference substance marked on detection antibody, from
And calculate the content of testing molecule.
The above embodiments are merely examples for clarifying the description, and does not limit the embodiments.For institute
For the those of ordinary skill in category field, other various forms of variations or change can also be made on the basis of the above description
It is dynamic.There is no necessity and possibility to exhaust all the enbodiments.And obvious variation extended from this or change
It moves still within the protection scope of the invention.
Claims (11)
1. a kind of detection method based on fluorescence-encoded magnetic bead, which is characterized in that include the following steps
Capture step: fluorescence-encoded magnetic bead is mixed with the product to be tested containing target molecule, captures target molecule；
It forms compound step: fluorescent reporter group is added, specificity is connected on captured target molecule, is formed fluorescence-encoded
Magnetic bead-target molecule-fluorescent reporter group compound；
Separating step: the fluorescence-encoded magnetic bead-target molecule-fluorescent reporter group compound is adsorbed using magnetic posts, and is turned
It moves on in reaction environment；
Excitation and determination step: it is compound that the fluorescence-encoded magnetic bead-target molecule-fluorescent reporter group is irradiated using exciting light
Object identifies the fluorescence-encoded magnetic bead-target molecule-fluorescent reporter group compound according to spectral waveform, according to glimmering
The fluorescence intensity of light reporter group carries out quantitative detection to captured target molecule.
2. the detection method according to claim 1 based on fluorescence-encoded magnetic bead, which is characterized in that the magnetic posts are column
Structure, magnetic posts working end diameter is 1mm-5cm, by current control magnetic state.
3. the detection method according to claim 1 or claim 2 based on fluorescence-encoded magnetic bead, which is characterized in that the reaction environment
Including buffer, confining liquid or sheath fluid.
4. the detection method based on fluorescence-encoded magnetic bead described in -3 any one according to claim 1, which is characterized in that further include sample
Product pre-treatment step is separated and is enriched with to target molecule in sample before capturing step.
5. the detection method based on fluorescence-encoded magnetic magnetic bead described in -4 any one according to claim 1, which is characterized in that further include
The preparation of fluorescence-encoded magnetic bead mixed liquor before capture step is mixed using at least two different fluorescence-encoded fluorescence-encoded magnetic beads
Carry out capture while at least two target molecules.
6. the detection method based on fluorescence-encoded magnetic bead described in -5 any one according to claim 1, which is characterized in that the capture
Step further include capture target molecule after elution and closing step and formed compound step in formed it is described fluorescence-encoded
Elution and closing step after magnetic bead-target molecule-fluorescent reporter group compound.
7. the detection method according to claim 6 based on fluorescence-encoded magnetic bead, which is characterized in that the elution and closing step
Suddenly it is adsorbed and is shifted using magnetic posts.
8. the detection method based on fluorescence-encoded magnetic bead described in -7 any one according to claim 1, which is characterized in that the excitation
Sepectrophotofluorometer, flow cytometer or liquid phase suspension array chip are selected from the detecting instrument that determination step uses.
9. a kind of detection kit using the detection method based on fluorescence-encoded magnetic bead described in claim any one of 1-8,
It is characterized in that, including,
It is coated with the fluorescence-encoded micro-beads of target molecule；
The detection antibody of fluorescent reporter group label；
Negative controls；
Positive reference substance；
Buffer.
10. detection kit according to claim 9, which is characterized in that fluorescence signal point on the fluorescence-encoded micro-beads
The sub and described fluorescent reporter group is selected from the different CdSe/ZnS quantum dot of center emission wavelength, the CdSe/ZnS amount
Sub- dot center's launch wavelength is 540-680nm.
11. detection method or claim 10 or 11 institutes described in a kind of any one of claim 1-8 based on fluorescence-encoded magnetic bead
Detection kit is stated in the application in fluorescence analysis field.
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Application publication date: 20190920 |