CN105510606A - Automatic immunoreactions detection apparatus - Google Patents
Automatic immunoreactions detection apparatus Download PDFInfo
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- CN105510606A CN105510606A CN201510422196.8A CN201510422196A CN105510606A CN 105510606 A CN105510606 A CN 105510606A CN 201510422196 A CN201510422196 A CN 201510422196A CN 105510606 A CN105510606 A CN 105510606A
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Abstract
An automatic immunoreactions detection apparatus relates to immunodetection, is provided with a reaction accelerating device and an integrated reagent mechanism; the reaction accelerating device is provided with an air pump, a guide pipe, a buffer flask, a gas-liquid guide pipe, an air throttle, a negative-pressure chamber and a pipe groove; the air pump is connected with the buffer flask via the guide pipe, the buffer flask is connected with the negative-pressure chamber via the gas-liquid guide pipe, the gas-liquid guide pipe is disposed at the bottom of the negative-pressure chamber, and the top of the negative-pressure chamber is provided with a pipe groove which is a concave cavity; the integrated reagent mechanism is disposed in the pipe groove; the integrated reagent mechanism is provided with an outer pipe, a sample-cell inner wall, a sample cell, an outer pipe bottom surrounding membrane fixed bulge, a combination membrane, a solid-phase membrane, a gasket, a liquid-guiding funnel and a flow-guiding pipe; the gasket is disposed on the outer pipe bottom surrounding membrane fixed bulge, the combination membrane and the solid-phase membrane are flatly disposed between the gasket and the sample cell, the combination membrane is disposed on the solid-phase membrane, the sample cell is inserted in the outer-pipe inner cavity, and the liquid-guiding funnel is disposed at the bottom of the outer pipe bottom surrounding membrane fixed bulge.
Description
Technical field
The present invention relates to immune detection, be specifically related to a kind of luminescence/fluorescence active immunity reaction checking device.
Background technology
Luminescence/fluoroimmunoassay technology is because of its high sensitivity, and good stability, and randomness operation in automation equipment, occupy core status, therefore instead of radiating immuning analysis technology rapidly in modern clinic immunological detection method.Immunologic function test reagent and use instrument must be highly merge, and competence exertion optimum performance, reaches best testing result.
The major measure of current optimization and accelerated reaction has: promote temperature to 37 DEG C, extends the reaction time, reactor concussion mixing, rotation mixing etc.Its objective is lifting molecular motion velocities, impel reactant fully to contact, to shorten the time that immune response reaches balance, thus ensure sufficiently high sensitivity.
Detect in analytical instrument work in active immunity, each step to be conducive to most the mode operation of detection reaction, all should to realize detection reaction and maximizes and optimization, to realize best Detection results.
Automatic mark immunoassay system comprises the multiple independent packaging components such as solid phase (micropore, microtubule, magnetic particle, plastic bead etc.), label, container, display system (colorimetric substrates, luminous substrate, fluorescence excitation light source), cleansing solution and runs instrument.The automatic detection system developed now relate to too many levels liquid transfer, and because of on Multi-example machine run, corresponding sample introduction, go out sampling device need repeatedly constantly clean, make instrument become huge, complicated, make difficulty, failure rate is higher; Only can detect the sample such as serum, blood plasma, can not whole blood sample be detected, be only limited to centralab and use; Longer because detecting turnover period (turnaroundtime, TAT), be not suitable for patient place Site Detection, be difficult to satisfied instant detection demand.
Denmark thunder degree RadiometerAQT90, Mitsubishi chemistry MitsubishiPATHFAST is all for whole blood sample Quantitative detection device.Other liquid components is distributed into the integrated morphology of single part by this kind of device in advance.This series products exist evaporate, storage life is short, Yi Jifen, encapsulate loaded down with trivial details and be easy to the problems such as cross pollution.And instrument and detection kind height single-minded, machine poor compatibility on sundry item, Testing index is limited, and extended capability is poor, is unfavorable for the utilization of resources.
The nitrocellulose filter solid phase side direction immunochromatography technique product being label with collaurum and fluorescence is that current one large class detects (PointofCareTest, POCT) product immediately.Easy is its outstanding feature.But because material variation is large, manufacture craft and testing process defectiveness, product quality poor controllability, physical strength is low.Make this series products sensitivity low, precision is low, poor repeatability, and dosing accuracy is not enough, can only be used for the primary dcreening operation to people at highest risk, can not be used as the conventional tool of medical diagnosis on disease.
Take cellulose acetate membrane as solid phase, collaurum is label spot immune percolation is also a kind of fast detection method.Its structure is that ad-hoc location point sample is fixed with specific antibodies (antigen) in cellulose acetate membrane, this immobilon-p and the absorbent material under it is fixed in a plastic casing.During detection, first upper sample, sample borrows capillary effect descending.After reaction certain hour, then instill colloid gold label thing, form solid phase-sample earnest-colloidal gold composite to be measured, have colour response thing, thus realize the qualitative detection to object.Unreacted composition is drawn in absorbent material under the capillary force action of absorbent material by cellulose acetate micropore.The method analysis ability is suitable with side direction immunochromatographiassays assays.Its solid phase procedures is extensive passive adsorption effect, and the amount of solid phase is uncertain, can not taken away determinand by the solid-phase component that adsorbs when subsequent detection, affect detection sensitivity; The fine and close light transmission of film is poor, and can hinder light transmition during luminesceence analysis, absorptiometric analysis is difficult to accurate quantitative analysis colorimetric, can only carry out qualitative or half-quantitative detection with scanning or ocular estimate; Colloid gold label thing can not be placed on solid phase NC film in advance, and detection needs to add colloid gold label thing in addition; Physical strength is low, can not fully wash, and reaction product is separated incomplete with unreacted reactant, efficiency is not high, also not thorough, therefore sensitivity is low; Can not the compositions such as separating red corpuscle be filtered, therefore can not be used for measuring whole blood sample.
Lateral chromatography immunoassay and spot immune diafiltration rule drive the mode that reactant liquor flows to carry out accelerated reaction process by capillary action.In a word, different detection methods and process, all need to take diverse ways to accelerate immunoreaction process.
Summary of the invention
In order to the sensitivity overcoming existing instant detection method (with collaurum and fluorescently-labeled side direction immunochromatography technique for representative) is low, precision is low and the problem such as poor accuracy, the invention provides a kind of active immunity reaction checking device, described active immunity reaction checking device comprises and can flow, accelerate the reaction accelerator of immune response speed and integrated reagent mechanism by accelerated reaction liquid.
Described active immunity reaction checking device comprises reaction accelerator and integrated reagent mechanism, and described reaction accelerator is provided with air pump, conduit, surge flask, gas-liquid conduit, air throttle, gas-liquid conduit, negative pressure chamber, tube seat; Described air pump and surge flask are by tubes connection, and described surge flask and negative pressure cavity are by gas-liquid tubes connection, and gas-liquid conduit is arranged at the bottom of negative pressure chamber, and described negative pressure chamber top is provided with tube seat, and described integrated reagent mechanism is arranged in tube seat; Described integrated reagent mechanism is provided with outer tube, sample cell inwall, sample cell, outer pipe bottom ring week film fixing prominent, binding film, immobilon-p, pad, drain funnel, mozzle; Described pad be arranged at outer pipe bottom ring week film fixing prominent on, described binding film, immobilon-p are flat between described pad and sample cell, and binding film is positioned on immobilon-p, and described sample cell is embedded in outer tube bore, described drain funnel is arranged at outer pipe bottom, and described mozzle connects drain funnel.
Described gas-liquid conduit is provided with air throttle.
Described tube seat is Reentrant Cavity.
Described outer tube is made up of plastic material, the hollow symmetrical structure of inside and outside sub-light, middle part is in funnel-form, scrub raffinate is derived outside pick-up unit by interstitial hole through this, and bottom is the mozzle of hollow, docks with the suction position of wash mill, by negative pressure, liquid waste is extracted out, complete washing, top is the fixing nest in fixed sample pond, and its interior bottom portion presses solidly mutually with sample pool side correspondence position determines film.
Described sample cell is made up of plastic material, for hollow, penetrates geometry up and down.
Described immobilon-p is the film having certain pore size, and aperture is not less than 20 μMs, and its effect is can homogeneous, immobilized antigen or antibody, filtering blood red blood cell, leucocyte expeditiously, and physical strength Gao Bingneng stands certain pressure liquid wash.
Described bond film is the film having certain pore size, and aperture is not less than 20 μMs, and its effect is storing marking antigen (antibody) bond, filtering blood red blood cell, leucocyte.
Described pad is poroid plastic sheet, plays support, protection immobilon-p and conjunctival effect during washing.
Described air pump is for generation of negative pressure, conduit is used for exhaust, and surge flask is used for compensator or trimmer pressure and stores waste liquid, and gas-liquid conduit is for discharging in negative pressure cavity the waste gas under taking out, waste liquid, air throttle is for controlling negative pressure cavity pressure, and negative pressure chamber is used for making reactant in integrated detector tube descending.Negative pressure locular wall is the closely knit plastic construction that can tolerate certain negative pressure, negative pressure chamber's upper edge and integrated testing agency sidewall close contact, avoids gas leakage.
The present invention compared with prior art, has following outstanding advantage:
The integrated reagent mechanism of the present invention is reasonably gathered in order by sample cell, film, pad, outer tube and is formed.The testing processes such as sample introduction, immune response, washing, luminescence-producing reaction and mensuration luminescence/fluorescence can be completed in this mechanism, eliminate the process of adding reagent label, solid formation, greatly shorten detection time, and have high sensitivity and good accuracy and repeatability.
Immune response accelerator of the present invention increases active immunity to detect in analytical instrument reagent flow in integrated reagent mechanism by arranging negative pressure cavity under integrated reagent mechanism, makes reactant fully contact and complete reaction.
The present invention can be used for immediately detecting accurate quantitative analysis; And can washed corpuscles and for the instant detection of whole blood sample.Be applicable to cells in sample particle, biomacromolecule and the micromolecular detections such as whole blood, serum, blood plasma, also for clinical patient sample is quick, sensitive, quantitative luminescence/fluoroimmunoassay is laid a good foundation.
Accompanying drawing explanation
Fig. 1 shows the structural representation of embodiment of the present invention active immunity detection reaction device;
Fig. 2 shows the structural representation of integrated reagent mechanism in embodiment of the present invention active immunity detection reaction device.
Embodiment
Embodiment 1
See Fig. 1 and Fig. 2, a kind of active immunity reaction checking device comprises reaction accelerator and integrated reagent mechanism 8, and described reaction accelerator is provided with air pump 1, conduit 2, surge flask 3, gas-liquid conduit 4, air throttle 5, gas-liquid conduit 4, negative pressure chamber 6, tube seat 7; Described air pump 1 is connected by conduit 2 with surge flask 3, described surge flask 3 is connected by gas-liquid conduit 4 with negative pressure chamber 6, and gas-liquid conduit 4 is arranged at the bottom of negative pressure chamber 6, and described negative pressure chamber 6 top is provided with tube seat 7, described tube seat 7 is Reentrant Cavity, and described integrated reagent mechanism 8 is arranged in tube seat 7; Described integrated reagent mechanism 8 is provided with outer tube 81, sample cell inwall 82, sample cell 83, outer pipe bottom ring week film fixing prominent 84, binding film 85, immobilon-p 86, pad 87, drain funnel 88, mozzle 89; Described pad 87 is arranged on all films of outer pipe bottom ring fixing prominent 84, described binding film 85, immobilon-p 86 are flat between described pad 87 and sample cell 83, binding film 85 is positioned on immobilon-p 86, described sample cell 83 is embedded in outer tube 81 inner chamber, described drain funnel 88 is arranged at bottom outer tube 81, and described mozzle 89 connects drain funnel 88.
Described gas-liquid conduit 4 is provided with air throttle 5.
Described sample cell inwall 82 is vertically parallel with outer tube 81, and interval 0.5 ~ 10mm, conveniently takes out sample cell.
Described outer tube 81 is made up of plastic material, the hollow symmetrical structure of inside and outside sub-light, middle part is in funnel-form, scrub raffinate is derived outside pick-up unit by interstitial hole through this, and bottom is the mozzle of hollow, docks with the suction position of wash mill, by negative pressure, liquid waste is extracted out, complete washing, top is the fixing nest in fixed sample pond, and its interior bottom portion presses solidly mutually with sample pool side correspondence position determines film.
Described sample cell 83 is made up of plastic material, for hollow, penetrates geometry up and down.
Described immobilon-p 86 is for there being the film of certain pore size, and aperture is not less than 20 μMs, and its effect is can homogeneous, immobilized antigen or antibody, filtering blood red blood cell, leucocyte expeditiously, and physical strength Gao Bingneng stands certain pressure liquid wash.
Described bond film 85 is for there being the film of certain pore size, and aperture is not less than 20 μMs, and its effect is storing marking antigen (antibody) bond, filtering blood red blood cell, leucocyte.
Described pad 87 is poroid plastic sheet, plays support, protects the effect of immobilon-p 86 and binding film 85 during washing.
Described air pump 1 is for generation of negative pressure, conduit 2 is for exhaust, and surge flask 3 is for compensator or trimmer pressure and store waste liquid, and gas-liquid conduit 4 is for discharging in negative pressure cavity the waste gas under taking out, waste liquid, air throttle 5 is for controlling negative pressure cavity pressure, and negative pressure chamber 6 is for making reactant in integrated detector tube descending.Negative pressure locular wall is the closely knit plastic construction that can tolerate certain negative pressure, negative pressure chamber's upper edge and integrated testing agency sidewall close contact, avoids gas leakage.
During detection, sample is added in sample cell 83, label aqueous phase dissolved in binding film 85, with the association reaction in film of the determinand in sample, booster air pump 1, the gas in conduit 2 pump drainage surge flask 3, in surge flask 3, negative pressure is ordered about in negative pressure chamber 6 via gas-liquid conduit 4 and is formed negative pressure under air throttle 5 flow control, order about reactant in integrated testing agency 8 descending gradually, when reactant is descending with solid formation association reaction on immobilon-p 86, formed solid phase-testing molecule-label reaction chain.After abundant reaction, sample liquid under certain strength effect, constantly wear film descending, new reaction is constantly formed, and makes sample liquid participate in reaction completely, forms solid phase-testing molecule-label reaction chain.
During washing, suitable for reading to sample cell injection washing lotion from sample cell, mozzle 89 docks and negative-pressure ward with vacuum line, and unreacted component is worn film and flow to drain funnel 88, is evacuated to outside pipe, completes washing by mozzle 89.During washing under certain descending strength effect, the educt (all compositions except tested composition) having neither part nor lot in reaction is drawn out of integrated reagent detector tube via fenestra is descending, completes washing, and it is residual to reduce washing lotion to greatest extent.Pad support and diaphragm play its function.
After having washed, in integrated reagent mechanism 8, add the luminous substrate liquid of certain volume, form film luminescence-producing reaction, measure luminous intensity (RLU) within a certain period of time, and got it right by the calibration object opisometer stored and answer testing concentration and report the result.
Embodiment 2: integrated reagent mechanism fluoroimmunoassay
As shown in Figure 2: pad 87 is positioned over bottom outer tube 81, respectively binding film 85, immobilon-p 86 are flat in outer tube 81 bottom gasket 87, then sample cell 83 is depressed in outer lumen, until binding film 85 and immobilon-p 86 are firmly fixed on all films of outer pipe bottom ring fixing prominent 84 by base, be combined into complete integrated testing agency.During detection, the sample of designated volume adds in sample cell, the label in binding film by aqueous phase dissolved, with the association reaction in film of the determinand in sample.Time descending with solid formation association reaction on immobilon-p, formed solid phase-testing molecule-label reaction chain.It is descending that the sample liquid of abundant reaction constantly wears film under certain strength effect, and new reaction is constantly formed, and makes sample liquid participate in reaction completely, form solid phase-testing molecule-label reaction chain.
During washing, suitable for reading to sample cell injection washing lotion from sample cell, outer pipe bottom mozzle 89 docks and negative-pressure ward with vacuum line, and unreacted component is worn film and flow to drain funnel 88, is evacuated to outside pipe, completes washing by outer pipe bottom mozzle 89.During washing under certain descending strength effect, the educt (all compositions except tested composition) having neither part nor lot in reaction is drawn out of integrated reagent detector tube via fenestra is descending, completes washing, and it is residual to reduce washing lotion to greatest extent.Pad support and diaphragm play its function.
After having washed, throw the exciting light according to certain wavelength to bottom binding film 85 and immobilon-p 86 at a certain angle from sample cell 83 side suitable for reading, and collect fluorescence intensity directly over film, got it right by the calibration object opisometer stored and answer testing concentration and report the result.
Claims (8)
1. an active immunity reaction checking device, it is characterized in that being provided with reaction accelerator and integrated reagent mechanism, described reaction accelerator is provided with air pump, conduit, surge flask, gas-liquid conduit, air throttle, gas-liquid conduit, negative pressure chamber, tube seat; Described air pump and surge flask are by tubes connection, and described surge flask and negative pressure cavity are by gas-liquid tubes connection, and gas-liquid conduit is arranged at the bottom of negative pressure cavity, and described negative pressure chamber top is provided with tube seat, and described integrated reagent mechanism is arranged in tube seat; Described integrated reagent mechanism is provided with outer tube, sample cell inwall, sample cell, outer pipe bottom ring week film fixing prominent, binding film, immobilon-p, pad, drain funnel, mozzle; Described pad be arranged at outer pipe bottom ring week film fixing prominent on, described binding film, immobilon-p are flat between described pad and sample cell, and binding film is positioned on immobilon-p, and described sample cell is embedded in outer tube bore, described drain funnel is arranged at outer pipe bottom, and described mozzle connects drain funnel.
2. active immunity reaction checking device as claimed in claim 1, is characterized in that described gas-liquid conduit is provided with air throttle.
3. active immunity reaction checking device as claimed in claim 1, is characterized in that described tube seat is Reentrant Cavity.
4. active immunity reaction checking device as claimed in claim 1, is characterized in that described outer tube is made up of plastic material, the hollow symmetrical structure of inside and outside sub-light.
5. active immunity reaction checking device as claimed in claim 1, is characterized in that described sample cell is made up of plastic material, for hollow, penetrates geometry up and down.
6. active immunity reaction checking device as claimed in claim 1, is characterized in that the aperture of described immobilon-p is more than 20 μMs.
7. active immunity reaction checking device as claimed in claim 1, is characterized in that the aperture of described bond film is more than 20 μMs.
8. active immunity reaction checking device as claimed in claim 1, is characterized in that described pad is poroid plastic sheet, plays support, protection immobilon-p and conjunctival effect during washing.
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| CN201510422196.8A CN105510606B (en) | 2015-07-17 | 2015-07-17 | A kind of active immunity reaction checking device |
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| CN201510422196.8A CN105510606B (en) | 2015-07-17 | 2015-07-17 | A kind of active immunity reaction checking device |
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| CN105510606A true CN105510606A (en) | 2016-04-20 |
| CN105510606B CN105510606B (en) | 2018-11-13 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110376373A (en) * | 2019-08-23 | 2019-10-25 | 王爱芳 | A kind of colloidal gold accelerates reaction unit and detection method |
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| EP0568664B1 (en) * | 1991-10-25 | 1998-07-15 | La Mina Ltd. | Capillary blood antigen testing apparatus |
| CN1553193A (en) * | 2003-06-06 | 2004-12-08 | 卢氏实验公司 | Counter colloidal gold immune chromographic reagent strips and their use |
| US20070202611A1 (en) * | 2003-12-24 | 2007-08-30 | Denka Seiken Co., Ltd. | Simple membrane assay method and kit |
| CN101201352A (en) * | 2006-12-13 | 2008-06-18 | 希森美康株式会社 | Instrument for forming solid phase of protein, protein solid phase forming device, and protein expression amount and activity value measuring device |
| CN204188614U (en) * | 2014-09-28 | 2015-03-04 | 南京钟鼎生物技术有限公司 | A kind of efficient immunoblotting reaction device |
| CN204807563U (en) * | 2015-07-17 | 2015-11-25 | 厦门先明生物技术有限公司 | Automatic immune response detection device |
-
2015
- 2015-07-17 CN CN201510422196.8A patent/CN105510606B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0568664B1 (en) * | 1991-10-25 | 1998-07-15 | La Mina Ltd. | Capillary blood antigen testing apparatus |
| CN1553193A (en) * | 2003-06-06 | 2004-12-08 | 卢氏实验公司 | Counter colloidal gold immune chromographic reagent strips and their use |
| US20070202611A1 (en) * | 2003-12-24 | 2007-08-30 | Denka Seiken Co., Ltd. | Simple membrane assay method and kit |
| CN101201352A (en) * | 2006-12-13 | 2008-06-18 | 希森美康株式会社 | Instrument for forming solid phase of protein, protein solid phase forming device, and protein expression amount and activity value measuring device |
| CN204188614U (en) * | 2014-09-28 | 2015-03-04 | 南京钟鼎生物技术有限公司 | A kind of efficient immunoblotting reaction device |
| CN204807563U (en) * | 2015-07-17 | 2015-11-25 | 厦门先明生物技术有限公司 | Automatic immune response detection device |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110376373A (en) * | 2019-08-23 | 2019-10-25 | 王爱芳 | A kind of colloidal gold accelerates reaction unit and detection method |
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