CN105435868B - Quantitatively detect the magnetic microparticle chemiluminescence micro-fluidic chip of Troponin I in whole blood - Google Patents
Quantitatively detect the magnetic microparticle chemiluminescence micro-fluidic chip of Troponin I in whole blood Download PDFInfo
- Publication number
- CN105435868B CN105435868B CN201510696728.7A CN201510696728A CN105435868B CN 105435868 B CN105435868 B CN 105435868B CN 201510696728 A CN201510696728 A CN 201510696728A CN 105435868 B CN105435868 B CN 105435868B
- Authority
- CN
- China
- Prior art keywords
- storage pool
- antibody
- substrate liquid
- magnetic particle
- micro
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of magnetic microparticle chemiluminescence micro-fluidic chip for quantitatively detecting Troponin I in whole blood; the micro-fluidic chip includes top plate (1) structure and bottom plate (2) structure, and the air pump (3), adding mouth (4), sample fill area (12), labelled antibody storage pool (5) and sample mixed zone (13) wherein on top plate (1) are sequentially connected；Filtering area (6), magnetic particle coating area (7), cleaning area (14), detection zone (8), liquid release channel (16) on bottom plate are sequentially connected；The detection zone (8) of bottom plate is connected with cleaning fluid storage pool (9) and luminous substrate liquid storage pool (10) by liquid release channel (16) respectively.
Description
Technical field
Realized the present invention relates to one kind using magnetic microparticle chemiluminescence technology and microfluidic chip technology in whole blood sample
The highly sensitive methods quantitatively detected of cTnI, particularly disclose a kind of magnetic particle chemistry hair for quantitatively detecting Troponin I in whole blood
Light micro-fluidic chip, accurate, the highly sensitive quantitative detection of cTnI in whole blood sample can be achieved, belong to fluidic chip chemiluminescence
Technical field of immunoassay.
Background technology
The prevention and control situation of current China angiocardiopathy is still severe, and cardiovascular disease incidence rate is in constantly rising situation.
According to statistics, cardiovascular death rate accounts for the 40% of human mortality, therefore carries out early discovery, early prevention, early treatment, and raising is cardiovascular
Sick preventing and treating level is crucial.Angiocardiopathy traditional detection project is mostly myocardial enzymes.But exist enzymatic activity rise occur compared with
The deficiencies of evening, short specific poor and duration.And cardiac troponin is the contractile protein for being uniquely present in cardiac muscle, to the heart
Myonecrosis has high susceptibility and specificity.
Cardiac troponin is by tri- kinds of cTnI, cTnC and cTnT, and important regulation is played in muscle diastole and contraction process
Effect.But cTnC is generally not used for myocardial damage detection without Cardiac-specific.CTnI and cTnT can not be penetrated under normal condition
Cell membrane enters blood, so cTnI and cTnT are extremely low in Healthy People blood；Such as myocardial cell damage, cTnI and cTnT enter people's cell
Interstitial and blood.In the diseases such as kidney failure, pneumonia and septicemia, cTnT contents can also raise in blood, so it is specific
Not as cTnI.CTnI is raised for 3~5 hours after morbidity, reaches peak within 15~24 hours, the duration is long, can be down to after 5~10 days
Normally.CTnI is one of myocardial injury markers best at present.
Traditionally use the detection such as ELISA, chemoluminescence method and immunochromatographic method (test strips) cTnI.It is but enzyme-linked
Immunization complex operation, detection time length；Chemoluminescence method needs supporting Large expensive instrument, testing time length, is not easy to realize soon
Fast detection immediately.Though colloidal gold immunity chromatography is easy to be quick, poor repeatability, sensitivity are low, easily judge by accident.
Chinese patent 200780015772.0 discloses a kind of troponin High Sensitive Analysis system, using microtiter plate
(microwell plate) is detected, high sensitivity, but complex operation, the testing time is long, detection range is narrow.Chinese patent
200610028913.X describes a kind of kit realized with colloidal gold immunochromatographimethod technology to cTnI detections, but can only carry out
Qualitative detection, and without standard measure.Chinese patent 201010619731.6 discloses a kind of whole immune layer for quantitatively detecting cTn I
Test strips are analysed, collaurum is substituted with fluorescent latex particles, realizes the quantitative detection to cTnI, but still can not solve test strips repetition
The defects of property difference.
Therefore quick, the accurate, detection method of high sensitivity is developed, there is great potential and application prospect.With it is glimmering
Light is compared with light is absorbed, and chemiluminescence does not have external excitation source background signal to disturb, and cross jamming is small, high sensitivity, linear
Scope is wide.The basic operation units such as sample preparation, reaction, separation, detection are integrated into one piece of micron meter by microfluidic chip technology
On the chip of degree, whole process analysis can be completed.
For the deficiency and defect of existing cTnI detection methods, micro-fluidic magnetic microparticle chemiluminescence method utilizes magnetic particle
Luminous and microflow control technique is learned, accurate to cTnI, highly sensitive quantitative detection can be achieved.
The content of the invention
The technical problem to be solved in the present invention is for existing fast diagnosis method sensitivity is low, poor repeatability, is disturbed
Substantially, and existing chemiluminescence necessary instrument is expensive, the problem of detection time is long, there is provided a kind of quantitatively to detect flesh calcium in whole blood
The magnetic microparticle chemiluminescence micro-fluidic chip of protein I, by integrated chip (in addition to test sample all components be integrated into
In chip) and supporting small portable device, so as to realize quick, accurate, the highly sensitive quantitative inspection of cTnI in live whole blood sample
Survey.
In order to solve the above technical problems, technical scheme provided by the invention is：
A kind of magnetic microparticle chemiluminescence micro-fluidic chip for quantitatively detecting Troponin I in whole blood, it is characterised in that described
Micro-fluidic chip includes top plate (1) structure and bottom plate (2) structure, and air pump (3), adding mouth (4), sample are filled out wherein on top plate (1)
Area (12), labelled antibody storage pool (5) and sample mixed zone (13) are filled to be sequentially connected；Filtering area (6), magnetic particle coating on bottom plate
Area (7), cleaning area (14), detection zone (8), liquid release channel (16) are sequentially connected；The detection zone (8) of bottom plate respectively with cleaning
Liquid storage pool (9) and luminous substrate liquid storage pool (10) are connected by liquid release channel (16)；
The labelled antibody storage pool (5) stores pre-packaged enzyme or luminous agent and marks anti-cTnI antibody, magnetic particle coating area
(7) the pre-packaged anti-cTnI antibody of magnetic particle marker is coated with, cleaning fluid storage pool (9) and luminous substrate liquid storage pool (10) storage are pre-
Encapsulate cleaning fluid and luminous substrate liquid；In the micro-fluidic chip testing process, the movement of magnetic particle or aggregation are manipulated with magnet；Institute
It is hydraulic seal pond to state labelled antibody storage pool, cleaning fluid storage pool and luminous substrate liquid storage pool, can by external force extrude and
Partial fracture, discharge liquid；The filtering area includes top plate (1) described in hemofiltration film and sealed with bottom plate (2) with adhesive tape (19 and 20).
Specifically, micro-fluidic chip of the present invention, luminous substrate liquid shelf-life should separate when being less than 1 year, with luminous
Substrate liquid storage pool A (23) and luminous substrate liquid storage pool B (24) substitutes luminous substrate liquid storage pool (10), the luminous substrate
Liquid storage pool A (23) and luminous substrate liquid storage pool B (24) is connected by pre-mixing passages (25).
Specifically, magnetic particle used in the present invention includes iron, cobalt, the compound of nickel, mainly including but not limited to three oxidations two
Iron and ferroso-ferric oxide compound.It is preferred that it is shell that magnetic particle, which is polystyrene, di-iron trioxide is the particle of core, magnetic particle size
There is obvious influence on testing result with the magnetic induction intensity of magnet.
Specifically, the magnetic particle size that the anti-cTnI antibody of the magnetic particle marker uses is 0.1~10 μm；Matched with magnetic bead
Magnet magnetic induction intensity be 500~30000 Gausses.
Preferably, the magnetic particle size that the anti-cTnI antibody of the magnetic particle marker uses is 0.5~3 μm, is matched with magnetic bead
Magnet magnetic induction intensity be 1000~8000 Gausses.
Specifically, enzyme or luminous agent mark cTnI antibody-solutions, magnetic particle marker cTnI antibody-solutions and the cleaning fluid
Buffer solution, protein, surfactant and preservative are included, and luminous agent mark cTnI antibody-solutions also include glycerine, magnetic
Particle marker cTnI antibody-solutions also include carbohydrate.
Specifically, enzyme mark cTnI antibody-solutions include bovine serum albumin(BSA) (BSA), Tween-20 and
Proclin300 borate buffer；Magnetic particle marker cTnI antibody-solutions include BSA, glucose, Tween-20 and
Proclin300 borate buffer；The cleaning fluid includes BSA, Tween-20 and Proclin300 borate buffer.
Specifically, the luminous agent mark cTnI antibody-solutions include BSA, glycerine, Tween-20, triton x-100 and folded
The phosphate buffer of nitrogen sodium；The magnetic particle marker cTnI antibody-solutions include BSA, casein, sucrose, Tween-20, Qula
Logical X-100 and Sodium azide phosphate buffer；The cleaning fluid include BSA, polysorbas20, triton x-100, polyethylene glycol and
The phosphate buffer of Sodium azide.
Specifically, the luminous substrate liquid includes substrate corresponding with enzyme and luminescence enhancement liquid, is injected after can merging luminous
Substrate liquid storage pool (10), or it is injected separately into luminous substrate liquid storage pool A (23) and luminous substrate liquid storage pool B (24).
Specifically, the luminous substrate liquid includes hydrogen peroxide solution and alkaline solution corresponding to luminous agent, is noted after can merging
Enter luminous substrate liquid storage pool (10), or be injected separately into luminous substrate liquid storage pool A (23) and luminous substrate liquid storage pool B
(24)。
Micro-flow control chip preparation method of the present invention is as follows：
Step 1) enzyme or luminous agent mark anti-cTnI antibody, and the anti-cTnI antibody of magnetic particle marker, both antibody can be identical
It is or different；
Enzyme or luminous agent labelled antibody solution are put into the labelled antibody storage pool of top plate by step 2), sealing, by magnetic
Grain labelled antibody solution is put into the coating area of bottom plate, is dried, and cleaning fluid and luminous substrate liquid are injected separately into cleaning fluid storage
In pond and luminous substrate liquid storage pool, sealing, top plate and bottom plate are assembled into micro-fluidic chip.
A kind of magnetic microparticle chemiluminescence micro-fluidic chip for quantitatively detecting Troponin I in whole blood provided by the invention is one
Kind realizes the micro-fluidic chip of quick, accurate, the highly sensitive detections of cTnI based on chemiluminescence, on micro-fluidic chip.
This chip is to make anti-cTnI antibody modifications enzyme, anti-cTnI antibody modifications using antigen-antibody on magnetic particle
With as whether contained cTnI in double antibody sandwich method principle combination magnetic particle rich, chemiluminescence detection whole blood sample, and accurately
Analyze its content.
Heretofore described enzyme, including but not limited to catalase (HRP) and alkaline phosphatase (ALP).Luminous substrate
Liquid is luminous substrate corresponding to enzyme (such as luminol or adamantane) and luminescence enhancement liquid (such as benzene derivative reinforcing agent), wherein sending out
Light substrate and luminescence enhancement liquid can merge, and inject a luminous substrate liquid storage pool (10) after being well mixed as shown in Figure 1；But work as
The mixed liquor shelf-life should separate when being less than 1 year, be injected separately into luminous substrate liquid storage pool A (23) and luminous substrate as shown in Figure 3
Liquid storage pool B (24), as shown in figure 3, it is well mixed by pre-mixing passages (25) during test, then flow into detection zone and participate in instead
Should.One embodiment of the invention uses horseradish peroxidase (HRP).
Luminous agent of the present invention, including but not limited to acridinium ester and acridine sulfonamide.Luminous agent is made with luminous substrate liquid
With rear, the catalytic action of enzyme is not required to, directly participates in luminescence-producing reaction.One embodiment of the present of invention uses acridinium ester.Luminous substrate
Liquid includes H2O2Solution and alkaline solution, alkaline H can be merged into2O2Solution, injection luminous substrate liquid storage pool (10)；But mix
When unstable afterwards, hydrogen peroxide solution and alkaline solution should be injected separately into luminous substrate liquid storage pool A (23) and luminous substrate liquid is deposited
Reservoir B (24), it is well mixed as shown in figure 3, first passing through pre-mixing passages (25) during test, then flows into detection zone and participate in reaction.
The cTnI antibody of the present invention includes monoclonal antibody and polyclonal antibody.The antibody can be combined (such as dual anti-with cTnI
Body sandwich method).Wherein the antibody of enzyme or luminous agent mark can be with identical with the antibody of magnetic particle marker, can also be different.
The enzyme or luminous agent labelled antibody solution and magnetic particle marker antibody-solutions of the present invention is comprising buffer solution, albumen
Matter, surfactant and preservative, and magnetic particle marker antibody-solutions also include carbohydrate.Wherein HRP labelled antibodies, buffer system
In can not contain NaN3；ALP labelled antibodies, buffer system can not be Phosphoric Acids.
The top plate of micro-fluidic chip and the moulding material of bottom plate of the present invention is polymer, including but not limited to polyphenyl second
Alkene, polyvinyl chloride, polypropylene, epoxy resin etc..Two-sided tape can be replaced by two one-faced tapes.As shown in figure 1, top board structure
By air pump (3), adding mouth (4), labelled antibody storage pool (5), lid (11), sample fill area (12) and sample mixed zone (13)
Composition.Base arrangement is by filtering area (6), magnetic particle coating area (7), detection zone (8), cleaning fluid storage pool (9), luminous substrate liquid
Storage pool (10), cleaning area (14), waste liquid pool (15) and liquid release channel (16).As shown in Fig. 2 in luminous substrate liquid and clearly
Washing lotion stores pool area, and magnet slide rail region, and the resigning hole for needing to reserve storage pool and magnet slide rail on top plate (is respectively
17 and 18), the resigning hole (being respectively 21 and 22) when storage pool should be reserved on double adhesive tape and sample mixed liquor flows into filtering area,
The effect of resigning hole is to get out of the way certain region, does not disturb liquid flow path, or necessary instrument part to act on micro-fluidic chip
Path.
The storage pool of the present invention is hydraulic seal pond, and encapsulant used includes glass, plastics, rubber, aluminium foil and high resistant
Every film, wherein encapsulant can be that same material forms, or multiple material combines.Under physical impact, storage
Pond can partial fracture, so that the liquid of sealing is discharged.Wherein enzyme mark cTnI antibody storage pool, cleaning fluid storage pool, hair
Light substrate liquid storage pool can use identical or different material and method to make.In one embodiment of the invention, enzyme mark cTnI
Antibody storage pool, cleaning fluid storage pool, luminous substrate liquid storage pool are sealed to form using plastics and elastic caoutchouc.The present invention's
In another embodiment, enzyme mark cTnI antibody storage pool is sealed to form using plastics and elastic caoutchouc, and cleaning fluid storage pool, hair
Light substrate liquid storage pool is sealed to form using high-isolation film.
The filtering area of the present invention includes hemofiltration film, and wherein hemofiltration film can make liquid by physical pore size or biology/chemical reagent
Body separates with cell, realizes that blood plasma separates with red blood cell, and blood plasma flows to magnetic particle coating area, and red blood cell rests on hemofiltration film
On, so as to reduce interference of the red blood cell to result of the test.Wherein described biology/chemical reagent includes coagulant etc., can make red thin
Intercellular connects, and forms grumeleuse, increased in size, it is easier to is stopped by the network structure of hemofiltration film.
The micro-fluidic chip of the present invention, when luminous substrate liquid storage pool A (23) and luminous substrate liquid storage pool B being present
(24) luminous substrate liquid pre-mixing passages (25) should, be increased on bottom plate, the pre-mixing passages can be serpentine channel or tie up and down
Structure hybrid channel, as shown in Figure 3.
In one embodiment, labelled antibody storage pool (5) encloses HRP and marks anti-cTnI antibody, coating area coating magnetic
Grain marks anti-cTnI antibody (different from enzyme labelled antibody), and cTnI is detected with magnetic particle enzyme-catalyzed chemical luminescence method.Another embodiment
In, labelled antibody storage pool (5) encloses the anti-cTnI antibody of acridinium ester label, the coating area coating anti-cTnI antibody of magnetic particle marker
(different from acridinium ester label antibody), cTnI contents are detected with magnetic particle enzyme-catalyzed chemical luminescence method.
The cleaning fluid of the present invention, for cleaning magnetic particle, removes cTnI, enzyme marker and other shadows of non-specific adsorption
Ring the material of testing result.Cleaning fluid mainly includes buffer system, protein and surfactant, wherein buffer system include but
It is not limited to borate, phosphate, Tris-HCl and acetate etc..Cleaning fluid pH 6.0~10.0, when detection sample be strong acid or
During strong basicity, pH scopes can relax.Wherein protein is including but not limited to bovine serum albumin(BSA), casein etc..Wherein surface is lived
Property is including but not limited to may include polysorbas20, Tween 80, triton x-100, polyethylene glycol and PVP etc..
The sample volume of the present invention is in 10~500 μ l, preferably 20~100 μ l.Preferably, the injection volume in embodiment
For 50 μ l.
The micro-fluidic chip of the present invention is quick detection, and detection time should be less than 30 minutes, preferably, being adopted in embodiment
With 15 minutes.
The antibody instrument of the present invention includes the functions such as extruding air pump and storage pool, magnet movement, luminescent detection system, Ying Ke
Include pressurizing unit, magnet and mobile device, detecting system, control analysis module and software systems.
The magnetic microparticle chemiluminescence micro-fluidic chip that a kind of cardiac muscle troponin I quantitatively detects, it is characterised in that described micro-
The testing process of fluidic chip includes：
After sample is instilled adding mouth by step 1), close the lid, micro-fluidic chip is put into necessary instrument, enzyme or luminous agent
After labelled antibody release, air pump is well mixed sample and labelled antibody, is then injected into bottom plate filtering area, and the necessary instrument is
Small portable device, include the functions such as extruding air pump and storage pool, magnet movement, luminescent detection system；
Behind the filtered area of step 2) sample, coating area is reached, dissolves magnetic labeling antibody, fully magnet collects magnetic after reaction
Grain, storage pool release cleaning fluid, after magnetic particle cleaning, moves to detection zone, discharges luminous substrate liquid, the detection of instrument detecting system
Luminous signal intensity, and then realize cTnI quantitative detection.
The core of the present invention realizes object using magnetic microparticle chemiluminescence immunoassay technology in micro-fluidic chip
Quickly, high sensitivity, accurate quantitative analysis detection.
Microfluidic chip technology is bases such as biology, chemistry, the sample preparation of medical analysis process, reaction, separation, detections
This operating unit is integrated on the chip of one piece of micro-meter scale, is automatically performed analysis overall process.
The micro-fluidic chip of the present invention is by all reagent components (enzyme mark cTnI antibody, the magnetic particle mark needed for detection process
Note cTnI antibody, cleaning fluid, luminous substrate liquid etc.) integrate, be built into micro-fluidic chip, and designed by ingenious raceway groove,
Under the operation of necessary instrument, realize that the one-touch detection of micro-fluidic chip (need to only be can be achieved with detecting, without multiple by start button
Miscellaneous operation), whole blood separation, immune response, cleaning separation, chemiluminescence detection are realized, so as to avoid existing micro-fluidic chip
Middle structure design is simple, detection when complex operation the deficiencies of and defect.Serum can only be carried out by also overcoming traditional chemical light-emitting appearance
Or blood plasma detection, and the shortcomings that can not be detected to whole blood sample.
Because magnetic particle easily precipitates, traditional chemical light-emitting appearance uses manual mixing, and maintains magnetic particle with persistent oscillation
Suspended state, but the operation that is mixed of magnetic particle is difficult to realize in miniature portable instrument in micro-fluidic chip.
Magnetic particle is coated with, dried in micro-fluidic chip raceway groove by the present invention, and devises magnet active drive magnetic particle
(and traditional microfluidic chip is typically using fluid driving or electric drive), so that magnetic particle redissolves, and in micro-fluidic chip not
Immune response, cleaning are realized with region, are lighted.This design not only solve magnetic particle be applied to easily to precipitate during micro-fluidic chip,
The problems such as poor repeatability, more controllable immune response and physical cleaning are also achieved, improves sensitivity and repeatability.
Micro-fluidic chip necessary instrument contacts with micro-fluidic chip no liquid in the present invention, no part for needing to clean, keeps away
Having exempted from traditional giant chemical light-emitting appearance needs stirring or sample-adding, cleaning etc. to operate and caused cross jamming and pollution.
So the present invention is not simple superposition magnetic microparticle chemiluminescence technology and microfluidic chip technology, but pass through liquid
Seal Design, raceway groove design, integrate all chemical constituents needed for detection, are built into micro-fluidic chip, and with magnet actively
Driving, realizes one-touch magnetic microparticle chemiluminescence immune detection, so as to realize cTnI in whole blood in portable necessary instrument
Quickly, high sensitivity, accurate quantitative analysis detection.
Present invention can apply to the quantitative detection of cTnI in angiocardiopathy especially heart failure.
Main advantages of the present invention are as follows：
1) present invention uses chemiluminescence method, has the advantages of low background, high sensitivity, wide range of linearity.
2) present invention uses magnetic granule technology, has magnetic enrichment function, strengthens simultaneously amplified signal；And magnet can be utilized magnetic
Particle transport zone (such as by being coated with area-cleaning area-detection zone), reduces the influence of sample matrix.
3) present invention uses microfluidic chip technology, and sample is mixed, reacted, is separated and detection is integrated on chip, and
All reagent components needed for reaction are integrated on chip.
4) present invention is easy to operate, during detection, only need to add sample, close the lid, it is supporting that chip is put into miniature portable
In instrument.
5) necessary instrument of the present invention is miniature portable instrument, and instrument is only physically contacted with chip, and liquid is not in chip
With instrument contacts, instrument will not be polluted and produce cross jamming.
Brief description of the drawings
Fig. 1 is that brain natriuretic peptide quantitatively detects micro-fluidic chip body structural representation, wherein 1 is top plate, 2 be bottom plate, and 3 be gas
Pump, 4 be adding mouth, and 5 be labelled antibody storage pool, and 6 be filtering area, and 7 be that magnetic particle is coated with area, and 8 be detection zone, and 9 be cleaning fluid
Storage pool, 10 be luminous substrate liquid storage pool, and 11 be lid, and 12 be sample fill area, and 13 be sample mixed zone, and 14 be cleaning
Area, 15 be waste liquid pool, and 16 be liquid release channel, and 17 be luminous substrate liquid and cleaning fluid storage pool resigning hole (in top plate), 18
For magnet slide rail resigning hole.
Fig. 2 is the complete microfluidic chip structure schematic diagram that quantitatively detects of brain natriuretic peptide, wherein 1 is top plate, 2 be bottom plate, 19
It is one-faced tapes for two-sided tape, 20,21 be luminous substrate liquid and cleaning fluid storage pool resigning hole (in two-sided tape), and 22 be mixed
Close resigning hole when liquid stream enters filtering area.
Fig. 3 is the micro-fluidic chip base arrangement schematic diagram of double luminous substrate liquid, wherein 23 be luminous substrate liquid storage pool
A, 24 be luminous substrate liquid storage pool B, and 25 be pre-mixing passages.
Embodiment
The invention discloses a kind of cTnI micro-fluidic magnetic microparticle chemiluminescence methods quantitatively detected and its special micro-flow control
Chip, those skilled in the art can use for reference present disclosure, be suitably modified technological parameter realization.In particular, institute
Have similar replacement and change it is apparent to those skilled in the art, they are considered as being included in the present invention.
The method of the present invention and application are described by preferred embodiment, and related personnel can be not substantially being departed from the present invention
Hold, method described herein and application be modified or suitably changed with combining in spirit and scope, to realize and using this
Inventive technique.
In order that those skilled in the art more fully understands technical scheme, with reference to specific embodiment pair
The present invention is described in further detail.
Embodiment 1：Enzyme-catalyzed chemical luminescence determines cTnI
(1) antibody labeling
Take 50 μ g HRP to be dissolved in 1mL distilled water, add 10 μm of ol and newly match somebody with somebody NaIO4Solution, the reaction of room temperature lucifuge
After 20min, with 1mM pH4.4 sodium-acetate buffer dialysis purification solution.PH is adjusted to 9.0 with pH9.5 carbonate buffer solutions again,
Add the anti-cTnI monoclonal antibodies of 100 μ g, room temperature lucifuge reaction 2h.0.1mL 4mg/mL are added newly to match somebody with somebody NaBH4, mix and react 2h after 4 DEG C.
Above-mentioned solution is loaded into bag filter, dialysed with 0.15M pH7.4PBS, 4 DEG C overnight, obtains HRP mark cTnI antibody.
1mg magnetic particle (a diameter of 2 μm), 10 μ g EDC and 15 μ gNHS solution and 10 are added into pH7.4 phosphate buffers
The anti-cTnI monoclonal antibodies of~30 μ g (different from the antibody of HRP marks) solution, is well mixed and reacts 4h at room temperature, it is sweet to add 1mg
Propylhomoserin is closed.With magnet enriching and purifying, unreacted cTnI monoclonal antibodies are removed, obtain magnetic particle marker cTnI antibody.
(2) micro-fluidic chip assembles
PH7.4 boron containing 1%BSA, 0.2% polysorbas20 and 0.05%Proclin300 in HRP mark cTnI antibody-solutions
Acid buffer；Magnetic particle marker cTnI antibody-solutions are to include 0.5%BSA, 2% glucose, 1% Tween-20 and 0.05%
Proclin300 pH7.4 borate buffers.
HRP labeling antibody solution is put into top plate labelled antibody storage pool, sealed.Magnetic labeling antibody solution is put into bottom plate magnetic
In particle coating area, drying at room temperature.
Cleaning fluid is 0.3%BSA, 0.2% Tween-20 and 0.03%Proclin300 pH7.2 borate buffers.Will be clear
Washing lotion injects cleaning fluid storage pool.Luminous substrate liquid is divided into HRP substrates (hydrogen peroxide solution of luminol) and alkalescence enhancing liquid (benzene
The alkaline solution of derivative), it is injected separately into luminous substrate liquid storage pool A (23) and luminous substrate liquid storage pool B (24), it is close
Envelope.As shown in Figure 1, filtering area is glued in bottom plate.Then as shown in Figure 2, with one-faced tapes and two-sided tape, by top plate and bottom
Plate is assembled into micro-fluidic chip.It is fitted into aluminium foil bag, seals 4 ° of preservations.
(3) pattern detection
Make dilution with human normal plasma, cTnI standard items are diluted to following concentration：0pg/ml、50pg/ml、100pg/
Ml, 500pg/ml, 1ng/ml, 5ng/ml, 10ng/ml and 50ng/ml.
After 50 μ l samples are instilled into adding mouth, close the lid.Micro-fluidic chip is put into necessary instrument, and (magnet magnetic induction is strong
Spend for 6000 Gausses) in, instrument extrusion HRP mark monoclonal antibodies, and make sample and HRP mark monoclonal antibody to inject bottom plate mistake after being well mixed
Filter area.After sample filtering, microchannel is reached, and dissolves magnetic particle marker monoclonal antibody, magnet accelerates sample reaction, forms HRP marks
The sandwich structure of monoclonal antibody-cTnI antigens-magnetic particle marker monoclonal antibody, then magnet collect magnetic particle.Storage pool discharges cleaning fluid,
After magnetic particle cleaning, the release of luminous substrate liquid, instrument detecting system detection luminous signal intensity.Total detection time 15min.Often
Individual sample is determined 3 times with 3 micro-fluidic chips respectively, is averaged, and draws standard curve.
50 μ l whole blood samples are instilled into adding mouth, instrument detecting system detects luminous signal intensity in 15 minutes, according to mark
Directrix curve obtains cTnI concentration in sample.
Cleaning Principle is：After whole blood adds micro-fluidic chip, whole blood first mixes with HRP labelled antibodies, then filtered
Qu Hou, the blood plasma for being mixed with HRP labelled antibodies reach microchannel, blood plasma dissolving magnetic marker antibody.When containing cTnI in blood sample, then
Form the sandwich structure (double antibody sandwich method) of HRP labelled antibody-cTnI- magnetic particle marker antibody.It is once purged, then light
The effect of substrate liquid is lower luminous, instrument detecting system test luminous signal.The standard curve obtained according to necessary instrument, and then analyze
CTnI concentration in blood sample.CTnI contents are higher in sample, then luminous signal is stronger.
As a result show, its lowest detection is limited to 50pg/ml, minimum to be quantitatively limited to 200pg/ml, and quantitative detection range is
0.05~50ng/ml, linearly dependent coefficient R2＞ 0.99；In detection range, do not occur HOOK effects；And weighed in criticizing and between criticizing
Renaturation is respectively less than 10%.Reference can be provided for heart infarction heart failure medical diagnosis on disease.
Embodiment 2：Direct chemical luminescent detecting cTnI
(1) antibody labeling
The acridinium ester activated in right amount and the anti-cTnI monoclonal antibodies solution of 100 μ g are added into phosphate buffer, be well mixed after
3h is reacted at room temperature, adds the closing of 1mg glycine.Dialysis isolates and purifies, and obtains acridinium ester label cTnI antibody.
1mg magnetic particle (a diameter of 1 μm), 20 μ g EDC and 20 μ g are added into lml 10mM pH7.4 phosphate buffers
NHS solution and 30 μ g Streptavidins, it is well mixed and reacts 4h at room temperature, adds the closing of 1mg glycine.With magnet adsorption
Enrichment, removes unreacted Streptavidin, obtains magnetic particle marker Streptavidin.
The anti-cTnI monoclonal antibodies of 20 μ g are added in 10 μ L 0.25mg/mL Sulfo-NHS-LC-biotin solution, react 1h.
Ultrafiltration centrifugal purification, unreacted biotin is removed, obtain biotin-cTnI antibody.
By the interaction between Avidin-Biotin, anti-cTnI antibody is connected to magnetic particle surface, obtains magnetic particle
Mark cTnI antibody.Wherein the magnetic particle of Avidin mark and biotinylated antibody ratios are 1: 104~2: 105。
(2) micro-fluidic chip assembles
Containing 0.5%BSA, 1% glycerine, 0.2% polysorbas20,1% triton x-100 in acridinium ester label cTnI antibody-solutions
With the pH7.4 phosphate buffers of 0.1% Sodium azide；Magnetic particle marker cTnI antibody-solutions are to include 0.2%BSA, 0.2% junket
Albumen, 1% sucrose, 0.5% polysorbas20, the pH7.4 phosphate buffers of 0.5% triton x-100 and 0.1% Sodium azide.Will
Acridinium ester label antibody-solutions are put into top plate labelled antibody storage pool, sealing.Magnetic labeling antibody solution is put into bottom plate magnetic particle
It is coated with area, drying at room temperature.
Cleaning fluid is the pH7.2 phosphoric acid of 0.3%BSA, 0.5% polysorbas20,1% triton x-100 and 0.02% Sodium azide
Salt buffer.Cleaning fluid is injected into cleaning fluid storage pool.Luminous substrate liquid is divided into comprising hydrogen peroxide solution and alkaline solution, difference
Inject luminous substrate liquid storage pool A (23) and luminous substrate liquid storage pool B (24), sealing.As shown in Figure 1, by hemofiltration film glue into
In bottom plate filtering area, storage pool is built into bottom plate.Then as shown in Figure 2, with one-faced tapes and two-sided tape, by top plate and bottom
Plate is assembled into micro-fluidic chip.It is fitted into aluminium foil bag, seals 4 ° of preservations.
(3) pattern detection
Make dilution with human normal plasma, cTnI standard items are diluted to following concentration：0pg/ml、100pg/ml、
500pg/ml, 1ng/ml, 5ng/ml, 10ng/ml and 50ng/ml.
After 50 μ l samples are instilled into adding mouth, close the lid.Micro-fluidic chip is put into necessary instrument, and (magnet magnetic induction is strong
Spend for 4000 Gausses) in, instrument extrusion acridinium ester label monoclonal antibody, and make to inject after sample and acridinium ester label monoclonal antibody are well mixed
In bottom plate filtering area.After sample filtering, microchannel is reached, and dissolves magnetic particle marker monoclonal antibody, magnet accelerates sample reaction, is formed
The sandwich structure of acridinium ester label antibody-cTnI antigens-magnetic particle marker antibody, then magnet collect magnetic particle.Deposit
Reservoir discharges cleaning fluid, and after magnetic particle cleaning, light exciting liquid release, instrument detecting system detection luminous signal intensity.Always
Detection time 15min.Each sample is determined 3 times with 3 micro-fluidic chips respectively, is averaged, and draws standard curve.
50 μ l plasma samples are instilled into adding mouth, instrument detecting system detects luminous signal intensity in 15 minutes, according to mark
Directrix curve obtains cTnI concentration in sample.
Cleaning Principle is：After whole blood adds micro-fluidic chip, whole blood first mixes with acridinium ester label antibody, then passes through
After filtering area, the blood plasma for being mixed with acridinium ester label antibody reaches microchannel, blood plasma dissolving magnetic marker antibody.When containing in blood sample
CTnI, then form the sandwich structure (double antibody sandwich method) of acridinium ester label antibody-cTnI- magnetic particle marker antibody.Through clear
After washing, light exciting liquid release, produces direct chemiluminescence with acridinium ester effect after mixing, and the test of instrument detecting system is luminous
Signal.The standard curve obtained according to necessary instrument, and then analyze cTnI concentration in blood plasma.CTnI contents are higher in blood plasma, then
Luminous signal is stronger.
As a result show, its lowest detection is limited to 80pg/ml, minimum to be quantitatively limited to 300pg/ml, and quantitative detection range is
0.08~50ng/ml, linearly dependent coefficient R2＞ 0.99；In detection range, do not occur HOOK effects；And weighed in criticizing and between criticizing
Renaturation is respectively less than 10%.Reference can be provided for heart infarction heart failure medical diagnosis on disease.
Embodiment 3：Magnetic particle particle size is screened
Other experiment conditions are carried out referring to embodiment 2, magnetic particle size and magnet magnetic induction intensity according to following scheme.
Particle size is 0.1 μm, 0.5 μm, 1.0 μm, 2.0 μm, 2.4 μm, 3 μm, 10 μm.Magnet magnetic induction intensity is 500
Gauss, 1000 Gausses, 4000 Gausses, 8000 Gausses, 12000 Gausses, 30000 Gausses.Driven respectively with this six kinds of magnet respectively
The magnetic particle of seven kinds of sizes.
Experimental result is shown：When 0.1 μm of magnetic particle and 500 Gauss magnet combine, its lowest detection is limited to 500pg/ml, fixed
Amount detection range is 0.5~50ng/ml, linearly dependent coefficient R2＞ 0.95；Batch in batch between repeatability respectively less than 20%.I.e.：
Chemiluminescence signal is weaker, and sensitivity is not high, and repeatability is poor.
When 10 μm of magnetic particles and 30000 Gauss magnet combine, its lowest detection is limited to 400pg/ml, and quantitative detection range is
0.4~5ng/ml, linearly dependent coefficient R2＞ 0.95；Batch in batch between repeatability respectively less than 20%.I.e.：Negative sample signal compared with
High (cleaning is insufficient), the range of linearity is not wide.
0.5~3 μm of magnetic particle is and the magnet of 1000~8000 Gausses is combined in use, its minimum detection limit is respectively less than
150pg/ml, quantitative detection range can reach 0.15~50ng/ml, linearly dependent coefficient R2＞ 0.97；Repeated in batch and between criticizing
Property is respectively less than 12%.Meet the needs that reference is provided for clinical heart infarction heart failure medical diagnosis on disease.
According to result above, preferably 0.5~3 μm of magnetic particle size, magnet magnetic induction intensity preferably 1000~8000 Gausses；
More preferably 1.0~3 μm of magnetic particle size, the Gauss of magnet magnetic induction intensity 4000~8000.Can according to used in magnetic particle size, enter
One step determines magnet magnetic induction intensity.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of magnetic microparticle chemiluminescence micro-fluidic chip for quantitatively detecting Troponin I in whole blood, it is characterised in that described micro-
Fluidic chip includes top plate (1) structure and bottom plate (2) structure, and air pump (3), adding mouth (4), sample wherein on top plate (1) are filled out
Area (12), labelled antibody storage pool (5) and sample mixed zone (13) are filled to be sequentially connected；Filtering area (6), magnetic particle bag on bottom plate
It is sequentially connected by area (7), cleaning area (14), detection zone (8), liquid release channel (16)；The detection zone (8) of bottom plate respectively with clearly
Washing lotion storage pool (9) and luminous substrate liquid storage pool (10) are connected by liquid release channel (16)；
The labelled antibody storage pool (5) stores pre-packaged enzyme or luminous agent and marks anti-cTnI antibody, magnetic particle coating area (7) bag
By the anti-cTnI antibody of pre-packaged magnetic particle marker, cleaning fluid storage pool (9) and luminous substrate liquid storage pool (10) storage are pre-packaged
Cleaning fluid and luminous substrate liquid；In the micro-fluidic chip testing process, the movement of magnetic particle or aggregation are manipulated with magnet；The mark
Remember that antibody storage pool, cleaning fluid storage pool and luminous substrate liquid storage pool are hydraulic seal pond, extruded by external force and local broken
Split, discharge liquid；The filtering area includes hemofiltration film, and the top plate (1) is sealed with bottom plate (2) with adhesive tape (19 and 20).
2. micro-fluidic chip as claimed in claim 1, it is characterised in that the luminous substrate liquid shelf-life should separate when being less than 1 year,
Luminous substrate liquid storage pool (10), the hair are substituted with luminous substrate liquid storage pool A (23) and luminous substrate liquid storage pool B (24)
Light substrate liquid storage pool A (23) and luminous substrate liquid storage pool B (24) is connected by pre-mixing passages (25).
3. chip as claimed in claim 1, it is characterised in that the magnetic particle chi that the anti-cTnI antibody of magnetic particle marker uses
Very little is 0.1~10 μm；Magnet magnetic induction intensity with magnetic grain matches is 500~30000 Gausses.
4. chip as claimed in claim 3, it is characterised in that the magnetic particle chi that the anti-cTnI antibody of magnetic particle marker uses
Very little is 0.5~3 μm, and the magnet magnetic induction intensity with magnetic grain matches is 1000~8000 Gausses.
5. micro-fluidic chip as claimed in claim 1, it is characterised in that enzyme or luminous agent mark the cTnI antibody-solutions,
Magnetic particle marker cTnI antibody-solutions and cleaning fluid include buffer solution, protein, surfactant and preservative, and luminous agent
Mark cTnI antibody-solutions also include glycerine, and magnetic particle marker cTnI antibody-solutions also include carbohydrate.
6. the micro-fluidic chip as described in claim 1 or 5, it is characterised in that the enzyme mark cTnI antibody-solutions include ox
The borate buffer of seralbumin (BSA), Tween-20 and Proclin300；Magnetic particle marker cTnI antibody-solutions include
BSA, glucose, Tween-20 and Proclin300 borate buffer；The cleaning fluid include BSA, Tween-20 and
Proclin300 borate buffer.
7. micro-fluidic chip as claimed in claim 1, it is characterised in that the luminous agent mark cTnI antibody-solutions include
BSA, glycerine, Tween-20, the phosphate buffer of triton x-100 and Sodium azide；The magnetic particle marker cTnI antibody-solutions
Include the phosphate buffer of BSA, casein, sucrose, Tween-20, triton x-100 and Sodium azide；The cleaning fluid includes
BSA, polysorbas20, triton x-100, the phosphate buffer of polyethylene glycol and Sodium azide.
8. micro-fluidic chip as claimed in claim 1, it is characterised in that the luminous substrate liquid includes substrate corresponding with enzyme
And luminescence enhancement liquid, after merging inject luminous substrate liquid storage pool (10), or be injected separately into luminous substrate liquid storage pool A (23) and
Luminous substrate liquid storage pool B (24).
9. micro-fluidic chip as claimed in claim 1, it is characterised in that the luminous substrate liquid includes double corresponding to luminous agent
The oxygen aqueous solution and alkaline solution, luminous substrate liquid storage pool (10) is injected after merging, or be injected separately into luminous substrate liquid storage pool A
And luminous substrate liquid storage pool B (24) (23).
10. micro-fluidic chip as claimed in claim 1, it is characterised in that the testing process of the micro-fluidic chip includes：
After sample is instilled adding mouth by step 1), close the lid, micro-fluidic chip is put into necessary instrument, enzyme or luminous agent mark
After antibody release, air pump is well mixed sample and labelled antibody, is then injected into bottom plate filtering area, and the necessary instrument is small-sized
Portable equipment, include the functions such as extruding air pump and storage pool, magnet movement, luminescent detection system；
Behind the filtered area of step 2) sample, coating area is reached, dissolves magnetic labeling antibody, fully magnet collects magnetic particle after reaction, deposits
Reservoir discharges cleaning fluid, after magnetic particle cleaning, moves to detection zone, discharges luminous substrate liquid, and the detection of instrument detecting system is luminous
Signal intensity, and then realize cTnI quantitative detection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510696728.7A CN105435868B (en) | 2015-10-26 | 2015-10-26 | Quantitatively detect the magnetic microparticle chemiluminescence micro-fluidic chip of Troponin I in whole blood |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510696728.7A CN105435868B (en) | 2015-10-26 | 2015-10-26 | Quantitatively detect the magnetic microparticle chemiluminescence micro-fluidic chip of Troponin I in whole blood |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105435868A CN105435868A (en) | 2016-03-30 |
| CN105435868B true CN105435868B (en) | 2017-11-28 |
Family
ID=55546760
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510696728.7A Active CN105435868B (en) | 2015-10-26 | 2015-10-26 | Quantitatively detect the magnetic microparticle chemiluminescence micro-fluidic chip of Troponin I in whole blood |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105435868B (en) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109201128A (en) * | 2017-07-05 | 2019-01-15 | 京东方科技集团股份有限公司 | Micro-fluidic chip, chemiluminescence immunoassay system and analysis method |
| CN107643284B (en) * | 2017-09-01 | 2018-12-18 | 北京华科泰生物技术有限公司 | For detecting the micro-fluidic chemiluminescence detection system of the magnetic particle of myocardium enzyme series |
| CN107655878B (en) * | 2017-09-01 | 2019-03-15 | 北京华科泰生物技术股份有限公司 | For detecting the micro-fluidic chemiluminescence detection system of the magnetic particle of thyroid function |
| CN108519373B (en) * | 2018-04-27 | 2024-03-15 | 广州万孚生物技术股份有限公司 | Chemiluminescence micro-fluidic chip and analysis instrument comprising same |
| CN109765358A (en) * | 2019-01-18 | 2019-05-17 | 江苏医联生物科技有限公司 | The chemical treatment method of micro-fluidic chip application protein detection |
| CN110244036B (en) * | 2019-07-01 | 2024-04-12 | 北京乐普诊断科技股份有限公司 | Microfluidic detection chip and method for immunodetection |
| CN111999225A (en) * | 2019-12-19 | 2020-11-27 | 瑞芯智造(深圳)科技有限公司 | Method for detecting concentration of micro-nano particles |
| CN113058666B (en) * | 2021-03-17 | 2023-02-07 | 广西科技大学 | Micro-fluidic chip for detecting Alzheimer disease related protein NOD-like receptor 3 |
| CN114011480B (en) * | 2021-11-04 | 2023-04-07 | 上海速创诊断产品有限公司 | Electrochemiluminescence microfluidic detection chip and kit for protein detection |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102713640A (en) * | 2009-06-10 | 2012-10-03 | 辛温尼奥生物系统公司 | Sheath flow devices and methods |
| CN103097029A (en) * | 2010-09-07 | 2013-05-08 | 多检测技术有限公司 | Assay device and reader |
| CN103575882A (en) * | 2013-11-15 | 2014-02-12 | 司珂 | Whole-blood labeled immunoassay method and instant detection system |
| CN205323796U (en) * | 2015-10-26 | 2016-06-22 | 深圳华迈兴微医疗科技有限公司 | Troponin I's magnetic particle chemiluminescence micro -fluidic chip in quantitative determination whole blood |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7439014B2 (en) * | 2006-04-18 | 2008-10-21 | Advanced Liquid Logic, Inc. | Droplet-based surface modification and washing |
-
2015
- 2015-10-26 CN CN201510696728.7A patent/CN105435868B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102713640A (en) * | 2009-06-10 | 2012-10-03 | 辛温尼奥生物系统公司 | Sheath flow devices and methods |
| CN103097029A (en) * | 2010-09-07 | 2013-05-08 | 多检测技术有限公司 | Assay device and reader |
| CN103575882A (en) * | 2013-11-15 | 2014-02-12 | 司珂 | Whole-blood labeled immunoassay method and instant detection system |
| CN205323796U (en) * | 2015-10-26 | 2016-06-22 | 深圳华迈兴微医疗科技有限公司 | Troponin I's magnetic particle chemiluminescence micro -fluidic chip in quantitative determination whole blood |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105435868A (en) | 2016-03-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105435868B (en) | Quantitatively detect the magnetic microparticle chemiluminescence micro-fluidic chip of Troponin I in whole blood | |
| CN105233892B (en) | Magnetic microparticle chemiluminescence double layer micro fluidic chip for whole blood sample detection | |
| CN105259163B (en) | The direct chemiluminescence micro-fluidic chip of magnetic particle for whole blood sample detection | |
| CN105241871B (en) | A kind of magnetic microparticle chemiluminescence micro-fluidic chip for whole blood sample detection | |
| CN105259162B (en) | Quantitatively detect the magnetic microparticle chemiluminescence micro-fluidic chip of brain natriuretic peptide in whole blood | |
| CN105435867B (en) | Detect the magnetic microparticle chemiluminescence micro-fluidic chip of creatine kinase isozyme in whole blood | |
| CN105241870B (en) | Detect the magnetic microparticle chemiluminescence micro-fluidic chip of n-terminal natriuretic peptide in whole blood | |
| CN205650212U (en) | A double -deck micro -fluidic chip of magnetic particle chemiluminescence for whole blood sample test | |
| CN205449995U (en) | Magnetic particle chemiluminescence micro -fluidic chip | |
| US9151749B2 (en) | Assay device comprising bubble-forming means | |
| CN105203775B (en) | The magnetic microparticle chemiluminescence micro-fluidic chip that a kind of Procalcitonin is quantitatively detected | |
| CN109870582A (en) | A kind of more target magnetic immunochemiluminescence micro-fluidic chip detection platforms and method | |
| CN105214744A (en) | A kind of magnetic microparticle chemiluminescence micro-fluidic chip | |
| CN105195243B (en) | The magnetic microparticle chemiluminescence micro-fluidic chip that a kind of myoglobins is quantitatively detected | |
| CN101793899B (en) | Optical biosensor for detecting brain natriuretic peptide (BNP) and preparation method of reagent thereof | |
| CN205317674U (en) | A direct chemiluminescence micro -fluidic chip of magnetic particle for whole blood sample test | |
| WO2021068913A1 (en) | Magnetic particle luminescence micro-fluidic chip for multi-marker detection, and detection device | |
| CN205650214U (en) | D - dimer quantitative determination's magnetic particle chemiluminescence micro -fluidic chip | |
| US20220184619A1 (en) | Magnetic particle luminescent micro-fluidic chip for multi-marker detection and detection apparatus | |
| CN205323796U (en) | Troponin I's magnetic particle chemiluminescence micro -fluidic chip in quantitative determination whole blood | |
| CN205599183U9 (en) | Magnetic particle chemiluminescence micro-fluidic chip for detecting creatine kinase isozyme in whole blood | |
| WO2021068914A1 (en) | Magnetic particle light-emitting double-layer micro-fluidic chip and detection system | |
| CN205449806U (en) | A magnetic particle chemiluminescence micro -fluidic chip for whole blood sample test | |
| CN205484063U (en) | Magnetic particle chemiluminescence micro -fluidic chip of terminal brain sodium peptide of nitrogen in detection whole blood | |
| CN211374779U (en) | Multi-marker detection magnetic particle luminous micro-fluidic chip and detection device |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |