CN105229468A - Have variable control line for the band that detects fast and the diagnostic kit using this band - Google Patents
Have variable control line for the band that detects fast and the diagnostic kit using this band Download PDFInfo
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- CN105229468A CN105229468A CN201480020367.8A CN201480020367A CN105229468A CN 105229468 A CN105229468 A CN 105229468A CN 201480020367 A CN201480020367 A CN 201480020367A CN 105229468 A CN105229468 A CN 105229468A
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Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
- G01N33/54386—Analytical elements
- G01N33/54387—Immunochromatographic test strips
- G01N33/54388—Immunochromatographic test strips based on lateral flow
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
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Abstract
The present invention relates to have variable control line for the band detected fast, the diagnostic kit comprising the band for detecting fast and for the method by analyzing thing in same use qualitative and quantitative analysis sample.Because the band for detecting fast of the present invention has variable control line, so it is possible for overcoming the difficulty of described analysis thing in quantitative test due to hook effect and utilize described naked eyes to carry out quantitative test highly reliably together with qualitative analysis in simple and quick mode.In addition, there is effect as follows: the dynamic range of described analysis thing can be improved by the complementary effect of described detection line and described variable control lines.
Description
Technical field
The present invention relates to have variable control line for the band detected fast, the diagnostic kit comprising the band for detecting fast and for by using in this band qualitative and quantitative analysis sample the method analyzing thing.
Background technology
Immunochromatographic method, be called as method for quick, for carrying out by the antigen-antibody reaction in short-term the method that qualitative and quantitative analysis analyzes thing on a small quantity, and be applied to multiple field, such as medical field, agricultural, livestock breeding industry, food, military affairs, environment etc., together with for the diagnosis of various diseases or inspection.Usually, utilize the analysis bar comprising and react with target analyte with the reagent changed, or utilize the analytical equipment comprising plastic casing and installation analysis bar in this box to implement this immunochromatographic method.Fig. 1 is the cross sectional view of the conventional analysis bar for immunochromatographic method.As shown in Figure 1, conventional analysis bar is made up of following: for receiving the sample pad of fluid sample; Comprise the pad of bond, this bond is by being attached to part (such as antigen or antibody) by the mark (such as, gold grain) of naked eyes or the detectable signal of sensor being prepared producing; Detecting pad, this detecting pad defines the detection line of bonding agent (antibody or antigen) and the control line for determining this sample migration that are fixed with and are specifically bound to and analyze thing and/or this bond in this sample; With the absorption layer for finally receiving this fluid sample.These functional cushions are partly overlapping each other by describing being linked in sequence into of they above, and be arranged in continuously on Solid support.If employ the analysis bar be arranged in plastic casing, sample reception hole then for sample being incorporated into sample pad is formed in the upper part of this box, and the position that the bonding agent that the result view window for observing this testing result is formed in this detecting pad is fixed.In the immunochromatographic method utilizing this analysis bar, when being expelled in this sample pad by fluid sample, fluid sample is flowed along this pad and this detecting pad by capillary action, and is finally adsorbed by absorption layer.Such as, an antibody that can be combined with target antigen is fixed (detection line) into a line on film pad, and other antibody and mark (in such as gold grain) combine, and then allow this sample migration.When the migration of this pad crossed over by this sample, its part produces the compound of antigen-gold grain antibody, and catches this antigen-gold grain antibody at fixing detection line place to be formed " antibody fixing at detection line-antigen-gold grain antibody place ".Now, this detection line is dyed to redness due to gold grain.So, this bond be contained in pad also moves together with this fluid sample.If target analyte is present in this sample, then this bond is incorporated into the bonding agent (being commonly referred to " interlayer reaction ") be fixed on this detecting pad by this analysis thing, or this bond maybe this analysis thing be attached to this bonding agent (being commonly referred to " competitive reaction ") competitively.Based on this conception of species, this analysis bar is for utilizing naked eyes or sensor to detect in sample the existence analyzing thing.
Invention discloses
Technical matters
This previously described method for quick, namely utilizes the immunochromatographic method of Quick kit, is usually in the form of band, then longitudinally cuts it prepare this band by fixing independent concrete material along some horizontal lines on long diaphragm.Therefore, advantage is, it carries out a large amount of production is in a simple manner decoupled possible; In addition, compared with enzyme linked immunosorbent assay (ELISA) etc., do not need extra washing step or second level reaction, and this reaction time is as short as 10 minutes.Therefore, this immunochromatographic method is fabulous in convenience.But shortcoming is, its detection limit is relatively low, and the cause owing to detecting based on naked eyes, accurate quantitative test is difficult.For this reason, this routine immunization chromatography is generally used for qualitative analysis, and there is many restrictions in its quantitative test.
Ladies and gentlemen inventor has improved this immunochromatography bar in quantitative test as possible together with the reliability in qualitative analysis, and therefore, they have been found that by adding containing being equal to the controlled control line analyzing thing or its analog, then contrast the signal intensity between this detection line and this variable control line, quantitative test is possible.Therefore, the invention provides the band for detecting fast with controlled control line, this band may be used for utilizing naked eyes to carry out analyzing in sample the reliable quantitative test of height of thing together with its qualitative analysis.
The solution of problem
In one aspect, the invention provides for carrying out the quick band detected with pad and detecting pad, wherein this pad has and marks with the first part of the first scheduled volume of analyte response and the input that is attached to this first part, this first part and this be marked at sample migration before or link each other when sample migration to form bond, and this bond can move to detecting pad when sample migration
Wherein this detecting pad has detection line separated from one another and variable control line,
Wherein this detection line is fixed together with the Ligands with analyte response in this sample, and
Wherein this variable control line is fixed together with the 3rd part of the second scheduled volume, and the 3rd part is equal to and this analysis thing of this first ligand reaction or its analog, thus can react with not being attached in sample the naked bond analyzing thing.
In a preferred embodiment, feature for the band detected fast is, analyze thing in the part of the first part of this first scheduled volume and this sample to react, with thus formed compound, wherein this analysis thing is attached to the bond with this first part and this mark, and the amount of the compound formed increases along with the increase of analyte concentration in this sample, but the amount of this naked bond reduces.
In another preferred embodiment, feature for the band detected fast is, this compound with the first part, mark and analysis thing reacts with the Ligands of catching at this detection line place, be not attached to the analysis thing in this sample and this naked bond with the first part and mark reacts with the 3rd part of catching at this variable control line place, and whether the analysis thing can determined in this sample by the signal measured from this detection line and this variable control line is existed or its amount or above-mentioned both.
Still in another preferred embodiment, the feature for the band detected fast is, the signal intensity of this variable control line reduces along with the increase of analyte concentration in this sample.
Also in another preferred embodiment, feature for the band detected fast is, signal intensity from this variable control line remains constant level, until the concentration analyzing thing in this sample reaches M value, but when this concentration reduces gradually more than this signal intensity during M value, and finally converge to 0, wherein when all 3rd parts fixed with this second scheduled volume by with this naked bond react reach capacity M value for sample in analyze the Cmax of thing.
Also in another preferred embodiment, the feature for the band detected fast is, this detecting pad has the constant control line for determining sample migration further, and this constant control line is fixed together with reporter molecule.
In addition in another preferred embodiment, the feature for the band detected fast comprises:
Sample pad, is incorporated in this sample pad by the sample comprising analysis thing to be analyzed;
Pad, one end of this pad is connected to this sample pad;
Detecting pad, one end of this detecting pad is connected to the other end of this pad;
Absorption layer, one end of this absorption layer is connected to the other end of this detecting pad, provides the driving force that this sample transports from sample pad; With
Be placed on the Solid support on the downside of the band for detecting fast.
Also in another preferred embodiment, the feature for the band detected fast is that this support member is made up of the material being selected from lower group, and this group is made up of the following: nitrocellulose, nylon, PVDF (Kynoar), glass and plastics.
Also in another preferred embodiment, the feature for the band detected fast is that this first part, this Ligands and the 3rd part are protein, antigen, antibody, DNA, RNA, PNA or fit.
Also in another preferred embodiment, the feature for the band detected fast is that this is labeled as aurosol, emulsion particle, colored ps particle, enzyme, fluorescent pigment, conducting polymer, luminescent substance or magnetic-particle.
Also in another preferred embodiment, the band for detecting fast is immunochromatography bar.
On the other hand, the invention provides the diagnostic kit of band for detecting fast comprising and being additionally arranged on as mentioned above in box, wherein this kit has guide and ribbon support part in lower box, there is sample reception hole in upper box, and result view window is placed on the position corresponding to this detection line and this variable control line.
In a preferred embodiment, the feature of diagnostic kit is that this kit is equipped with standard signal data, and this standard signal packet is containing for containing the detection line of sample of analysis thing and the standard colorimetric table of variable control line that are in multiple concentration.
In another preferred embodiment, the feature of this diagnostic kit is whether can be existed and intensity by the signal of the mark with the naked eye checking detection line and variable control line, implement qualitative or semi-quantitative analysis.
Still on the other hand, the invention provides and utilize above-mentioned carrying out quantitatively or the method for qualitative analysis analyzing thing in sample for the band detected fast,
The method comprises the steps:
This sample is incorporated into this pad or in the pad of the prelocalization of this pad, and allows this sample migration (step 1);
Detect and whether exist and intensity (step 2) from the signal of this detection line with the mark of this variable control line; With
The signal intensity of this detection line and this variable control line and standard signal data are carried out the amount contrasting to determine this analysis thing,
Wherein by making each the experience step 1 and 2 in the sample of this analysis thing comprising different concentration known obtain this step signal data (step 3).
In a preferred embodiment, the method is characterized in that, in step 2, utilize opacimeter inspection whether to exist and signal intensity from the signal of this detection line with the mark of this variable control line.
Still on the other hand, the invention provides in sample, one or more analyze the method for the quantitative of thing or quantitative test, comprise and this sample and the part be adsorbed on Solid support are reacted, it is characterized in that analyzing thing to one is applied with two kinds of different reaction mechanisms.
In a preferred embodiment, the method is characterized in that these two kinds different reaction mechanisms are interlayer reaction and competitive reaction.
Favourable effect of the present invention
Because the band for detecting fast of the present invention has variable control line, so to analyze the difficulty of thing in quantitative test and utilize naked eyes to carry out quantitative test highly reliably together with qualitative analysis in simple and quick mode be possible because hook effect overcomes.In addition, there is effect as follows: the dynamic range analyzing thing can be improved by the complementary effect of detection line and variable control line.
Accompanying drawing explanation
Fig. 1 shows the schematic diagram of the viewgraph of cross-section for the analysis bar in typical immunochromatographic method;
Fig. 2 shows the schematic diagram for the analysis bar in immunochromatographic method of the present invention;
Fig. 3 shows before and after introducing sample, the schematic diagram of the pattern of detection line, variable control line and constant control line in the band for detecting fast of the present invention;
Fig. 4 illustrates the schematic diagram for the band detected fast according to one embodiment of present invention; With
Fig. 5 is the figure showing the result analyzing analyte concentration in several samples according to one embodiment of present invention, wherein human immunoglobulin G (IgG) thing that performs an analysis.
Implement best mode of the present invention
In one aspect, to achieve these goals, the invention provides the band for detecting fast with pad and detecting pad, wherein this pad has and marks with the first part of the first scheduled volume of analyte response and the input that is attached to this first part, this first part and this mark link each other with before sample migration or sample migration time form bond, this bond can migrate to detecting pad when sample migration, this detecting pad has detection line separated from one another and variable control line, this detection line is fixed with analyzing in sample together with Ligands that thing reacts, and the 3rd part of this variable control line and the second scheduled volume reacts, 3rd part is equal to the analysis thing or its analog that react with this first part, thus can react with not being attached in sample the naked bond analyzing thing.
The invention is characterized in, variable control line fixing together with being equal to the material of this analysis thing or its analog is added to the conventional analysis bar with detecting pad, this detecting pad only defines the detection line for guaranteeing this sample migration and control line, thus performs quantitative test highly reliably together with qualitative analysis.
In addition, the invention is characterized in, under the prerequisite not having specialty analysis instrument, with the naked eye perform semi-quantitative analysis highly is reliably possible.
In a preferred embodiment, the invention is characterized in that the band for detecting fast is immunochromatography bar.
As this is used, term " immunochromatographic method " be combine based on antigen-antibody reaction the analytical approach of immune response principle and the red, orange, green, blue, yellow (ROGBY) principle of being moved along medium by mobile phase based on sample and reagent.In brief, antibody to be analyzed or antigen to be pre-allocated on perforated membrane and to be fixed to the upper, and then allow blood from one end migration of this film to antibody or antigen to observe the reaction of antigen or antibody it and blood.Immune response typically refers to antigen-antibody reaction.But in the present invention, it also broadly comprises the reaction between acceptor and part that is bonded to each other specifically together with antigen-antibody reaction, but is not limited to this.In addition, this immune response is comprised and mutually being identified by specificity and responding of occurring, such as, reaction between enzyme and substrate.
Immunochromatographic method can occur in comprise analyzes the sample of thing when moving through medium by capillary action along mobile phase.Therefore, as the medium for this immunochromatographic method, can prepare and use band.Will hereinafter be described for the particular elements of band that detects fast and function thereof.
Can usage flag so that with the naked eye or sensor detectable antigens-antibody response simply.The part that can be attached to this analysis thing specifically can be linked to this mark, to make this mark be attached to target analytes.
As this is used, term " bond " refers to the bond by this mark and this part are linked each other and formed, and this bond and this label link for input and comprise with sample in analyze the first part that thing reacts.In order to distinguish the first part and other parts hereinafter described, this first part refers to the part be combined with this mark to form bond.
As used in this, this first part refers to and can react with this analysis thing and be attached to the material of this analysis thing specifically.
Physically or chemically can link this mark and this first part.That is, this mark and this first part can be linked to have reactive group via passive adsorption or covalent bond by changing mark, but be not limited thereto.The link can implementing between this mark and this first part by methods known in the art.
But, this mark and this first part can be present in this pad with the form of bond before the sample migration in the band for detecting fast, or do not having to be present in individually on this pad under the prerequisite linked, and then they are bonded to each other during sample migration, form this bond.No matter which kind of situation, this mark and this first part move to this detecting pad with the form of bond during sample migration, because they are not fixed on this pad.
As this is used, term " mark " refer to generation can with the naked eye or sensor the material of signal detected.This mark can be aurosol (gold grain), emulsion particle, coloured ps particle, enzyme, fluorescent pigment, conducting polymer, luminescent substance, magnetic-particle etc., but is not limited thereto.In addition, this signal can be the cold light by the spontaneous generation of the intrinsic property of this mark, or by fluorescence that outside stimulus produces.
As used in this, term " part " refers to the material be bonded to each other specifically.Such as, be attached to the antibody of antigen specifically or be attached to the part of concrete acceptor specifically, working using as part each other.The non-limitative example of this part comprises protein, antigen, antibody, DNA, RNA, PNA or fit.In addition, the part in the present invention can be any material and unrestricted, as long as it shows characteristic as defined above.Run through the part that this instructions uses and refer to the material be bonded to each other specifically, as above limit, unless be constrained to the part being attached to concrete acceptor specifically.
In a preferred embodiment of the invention, this mark and this first part are connected to each other in advance and are present on this pad with the form of bond.In this case, in the solution of concentration known, prepare this mark and this first part individually, then mix them and allow their reaction schedule times to prepare bond solution.This bond solution is distributed in the pad this pad comprising this bond with preparation.Now, consider the combining ratio of this mark and this part, concentration and the mixture ratio of each solution can be determined.The combination of this mark and this part can be one and mark the combination with a part, a mark and the combination of multiple part or the combination of multiple mark and a part.Specific binding ratio is unimportant, but it can keep estimated rate in a preferred embodiment.Combining ratio can change according to the type of this mark and this part, and can be determined by the amount of the relative size and binding site of considering them.Such as, if some micron order emulsion particles are used as this mark and antibody be used as this part, then multiple antibody can be incorporated into an emulsion particle.Preferably, in order to the antibody of equivalent being attached to each in latex surface, concentration and their mixture ratio of antibody and latex solution can be controlled, with make antibody and latex surface in conjunction with saturated, but to be not limited thereto.
As mentioned above, can by mixing this mark of concentration known and the first part and making them react to obtain the bond solution of concentration known.Bond solution can in the form of dispersion, and this bond molecule wherein with predetermined concentration is evenly dispersed in solution.
Band for detecting fast is the immunochromatography bar with variable control line, and this immunochromatography bar can comprise the pad containing and mark with the input being attached to this first part with the first part analyzing the first scheduled volume that thing reacts; And there is the detecting pad of detection line and variable control line individually.
To analyzing the quantitative test of thing together with qualitative analysis in sample, the 3rd part is fixed on this detecting pad to form " variable control line ".In the variable control line of the band for detecting fast of the present invention, the material being equal to analysis thing or its analog reacted with this first part can be fixed as the 3rd part, and the 3rd part can react with being in the naked bond that mobile phase flows in this detecting pad.The naked bond not being attached to the analysis thing in this sample comprises this first part, therefore, is attached to the material being equal to this analysis thing or its analog to this first ligand specificity, i.e. the 3rd part.
In addition, the 3rd part of scheduled volume can be fixed on this variable control line.That is, this second scheduled volume refers to the specified quantitative of the 3rd part be fixed on this variable control line.By fixing the 3rd part of this second scheduled volume, what can control to analyze in signal intensity in variable control line and this sample thing can detectable concentration scope, and this makes quantitative test highly reliably become possibility.
In the present invention, in order to distinguish the 3rd part and other parts hereinafter described, the 3rd part refers to and is fixed on this detecting pad to form the part of this variable control line.
In addition, in order to detect the analysis thing in this sample, be attached to this analysis thing specifically and the Ligands of optionally catching is fixed on this detecting pad, thus form " detection line ".In the detection line of the band for detecting fast of the present invention, and analyzing the Ligands that thing reacts in this sample and fix to react with analysis thing-bond compound (being namely in the analysis thing that mobile phase flows in this detecting pad).This Ligands can for being equal to this first part or its analog or being attached to the material of the 3rd material of this analysis thing specifically.When the analysis thing in this sample moves along mobile phase, it is attached to bond in this pad to form compound.Because this compound comprises as above the first part analyzed thing and be attached to this analysis thing specifically, contribute to catching compound in this detection line by being attached to this analysis thing so be equal to the first part or its analog or this material of Ligands of being combined with this analysis thing specifically.
In the present invention, in order to distinguish Ligands and other parts hereinafter described, this Ligands refers to and is fixed on this detecting pad to form the part of this detection line.
As mentioned above, this immunochromatographic method utilizes the red, orange, green, blue, yellow (ROGBY) based on the mobile phase migration comprising this analysis thing along medium.Therefore, in the immunoassay of the band for detecting fast used according to the invention, mobile phase is needs for the sample comprising this analysis thing for this band migration.Therefore, immunoassay kits of the present invention may further include damping fluid.Damping fluid serves as the mobile phase along this sample of bar Tape movement for detecting fast, and serves as the solution for dissolving this bond.If desired, it serves as the thinning agent for diluting this sample.Such as, with regard to whole blood, may further include for dissolving haemocyte, the composition of such as red blood cell etc.As damping fluid, the phosphate buffer (PBS) of typical damping fluid such as 10mM to 1M, nonionic or amphoteric surfactant or their potpourri can be used under hard-core prerequisite, and can according to required reaction, such as antigen-antibody reaction these, suitably select damping fluid.
This pad is that this first part with the first scheduled volume reacted with this analysis thing as described above marks with the input being attached to this first part, wherein this first part and this be marked at this sample migration before can the form of bond be present on this pad, or be present in individually under the prerequisite not having to link between them on this pad, then they are bonded to each other to form bond during sample migration.
In the present invention, only this first part of scheduled volume may reside in this pad.That is, this first scheduled volume refers to the specified quantitative of the first part be present in this pad.Because this first part is present in this pad with the first scheduled volume, so the amount being combined the compound formed by this first part with this target along with sample hit target concentration increase and increase, but the amount not being attached to the naked bond of target reduces, and is shown as inverse ratio.
Preferably, in this pad, this first part and this mark link in advance each other, and they exist with bond form.Now, this pad can be prepared by the bond of concentration known is applied to pad, as described above.
In addition, this pad may further include the second bond being combined by the 4th part and mark and formed, and waits to be used as internal contrast thing.4th part refers to the part that can react with reporter molecule specifically or non-specifically.Catch the second bond by the reporter molecule of constant control line, thus guarantee the migration of this sample.By in the detailed description hereafter carrying out this constant control line and this report molecule.
This detecting pad is the medium for mobile phase and sample migration, and this mobile phase and this sample can be moved by the capillary action forming the perforated membrane of this detecting pad.In addition, this detecting pad has detection line separated from one another and variable control line, and has the constant control line for guaranteeing this sample migration further.One end of this detecting pad can be connected to this pad, and its other end can be connected to this absorption layer, and this absorption layer is provided for the driving force of sample migration.This pad and this absorption layer partly overlap onto on this detecting pad.This detecting pad can be made up of perforated membrane, and this perforated membrane can be nitrocellulose membrane, glass fibre membrane, polyethersulfone (PES) film, cellulose membrane, nylon membrane or their combination, but is not limited thereto.Preferably, this detecting pad can for having the nitrocellulose membrane of 5 to 15 micron pore size.
To the principle utilizing the qualitative or quantitative test of the band for detecting fast of the present invention hereafter described in more detail.
In conventional analysis bar, this detection line is formed on this film pad, and the analysis thing-bond compound of catching this migration at this detection line place is to produce signal (interlayer reaction).Now, signal intensity increases along with the concentration increase of this analysis thing.But if the concentration of this analysis thing exceedes certain concentration, then signal intensity reduces due to hook effect.This phenomenon is the huge obstacle utilizing conventional analysis bar to carry out analyzing thing quantitative test.
But, the feature of the band for detecting fast of the present invention is, specific competitive reaction (analyze in the sample in this first part and limited amount pad of the first scheduled volume thing react competitively) is reacted together with interlayer and is applied to a band, therefore, compared to routine immunization chromatographic analysis, perform more fast, quantitative test is easily possible.
Specifically, when in sample pad fluid sample being incorporated into the band for detecting fast of the present invention, mobile phase and sample occur.In this pad, in this first part of the first scheduled volume and this sample, limited amount analysis thing reacts competitively.Analysis thing in a part for this first part of the first scheduled volume and sample reacts to form compound, wherein this analysis thing is attached to the bond comprising the first part and mark, but the remainder of this first part do not reacted with this analysis thing moves along mobile phase as naked bond.Herein, because this first part remains the first scheduled volume, the amount of all formed compounds increases along with the increase of analyte concentration in this sample, but the amount of this naked bond reduces.
The Ligands of catching at this detection line place during the compound comprising this first part, this mark and this analysis thing and migration reacts, but with this sample does not analyze thing and to react and the 3rd part of catching at this variable control line place during the naked bond comprising this first part and this mark and migration reacts.Respectively at this detection line and this variable control line place capture complexes and naked bond, because this Ligands is fixed on this detection line and the 3rd part is fixed on this variable control line.
This compound and this naked bond all comprise mark, therefore can by measuring from catching that this compound and this detection line of this naked bond and the signal of this variable control line determine whether this analysis thing exists, in this sample they amount or above-mentioned both.
As mentioned above, the amount of this compound increases along with the increase of analyte concentration in this sample, but the amount of this naked bond reduces along with the increase of analyte concentration in this sample.Therefore, the signal intensity from the variable control line of catching this naked bond reduces along with the concentration increase analyzing thing in this sample.
More specifically, the signal intensity from this variable control line remains on constant level until analyte concentration reaches M value in sample, but this signal intensity reduces more than M value gradually in this concentration.Finally, this signal intensity converges to 0.Herein, M value be when all 3rd parts fixed with the second scheduled volume by with this naked bond react reach capacity this sample in analyze the Cmax of thing.In an example of the present invention, when the concentration analyzing thing in this sample is in 0 to 90ng/ml scope, the signal intensity from this variable control line remains on the strongest level, and therefore all 3rd parts and this naked bond react as seen.Therefore, confirm that the M value in example of the present invention is about 90ng/ml (example 2).But M value changes according to the type of the second scheduled volume and the 3rd part and this analysis thing, therefore, M value can suitably be selected by those skilled in the art according to the object analyzed.
As mentioned above, the signal intensity from this detection line increases along with the increase of this analyte concentration.But the concentration of this analysis thing exceedes specified level, signal intensity reduces due to hook effect.According to the concentration that analysis thing of the present invention increases, the signal intensity figure of this detection line and this variable control line is shown in Figure 3.
Therefore, the qualitative of this analysis thing is performed by the signal measured from the mark of this detection line and this variable control line or quantitative test is possible.More particularly, from the signal of this detection line and this variable control line and the standard signal Data Comparison of the analysis thing of previously prepared concentration known, thus the concentration of this analysis thing is analyzed.This concentration of semi-quantitative analysis can be carried out by utilizing naked eyes to detect this signal intensity, or utilize reader such as opacimeter more accurately this concentration of quantitative test.
In the present invention, this detecting pad has the constant control line for guaranteeing this sample migration further, and this report molecule can be fixed on this constant control line.
As this is used, term " constant control line " refers to the concentration not considering to analyze thing in this sample or this sample and produces the part of constant signal.Mode that can be similar with this variable control line to forming this detection line forms this constant control line.But, this constant control line can be formed by fixed ligands thereon, wherein said part is not attached to this target material (analysis thing), and specifically or be non-specifically attached to and catch the 4th part of this second bond moved by mobile phase along this detecting pad together with this sample.Alternately, this constant control line can be formed by fixed ligands thereon, wherein said ligand specificity ground or be non-specifically attached to and catch the mark that moved by mobile phase along this detecting pad together with this sample or mark-in conjunction with material.Therefore, as this part, do not considering that the material that can produce constant signal under analyte concentration in this sample and the prerequisite that whether exists is fixed, to form this constant control line.The part being ready to use in this constant control line is called term " reporter molecule ", and the example of this report molecule can comprise anti-rabbit IgG, anti-chicken IgY, Streptavidin, bovine serum albumin(BSA) etc.
Two or more in this constant control line are formed by the concentration changing reporter molecule to be fixed thereon.When the concentration being fixed on the reporter molecule on this constant control line increases, this constant control line can produce strong signal consistently.
If this report molecule of concrete concentration is fixed on this constant control line, then consequent signal intensity remains on constant level, therefore can determine to be in the concentration of the analysis thing corresponding to this detection line of this report molecular signal intensity or the signal intensity of this variable control line.Some constant control lines can be formed by the concentration changing this report molecule, thus form the some constant control line with unlike signal intensity.For quantitative test compared with the signal intensity of these signal intensities and this detection line or this variable control line.That is, this constant control line can be used as internal standard signal data.
Such as, suppose to define two constant control lines, and they produce the signal intensity of 1 and 3.Based on the analysis thing of concentration known, can pre-determine, when this detection line has signal intensity 1, the concentration of this analysis thing is 10ng/ml, and when this detection line has signal intensity 3, the concentration of this analysis thing is 30ng/ml.If this detection line display intensity 2 (deriving from the analysis that unknown concentration analyzes thing), then can find out that this number intensity is the signal intensity of the intermediate value (signal intensity compared to this constant control line) corresponded between two constant control lines simply.
Correctly can select size and the position of this detection line, this variable control line and this constant control line according to antigen-antibody reaction to be used, but be not limited thereto.The Successful migration determining this sample whether can be there is by the signal from this constant control line, and can by the signal intensity of this detection line and this variable control line and the standard signal Data Comparison previously determined being implemented the quantitative test of this analysis thing.Will be described below standard signal data.
In the present invention, when the signal intensity by this detection line and this variable control line carries out quantitative test, the dynamic range analyzing thing in this sample can be 1ng/ml to 1mg/ml.In an example of the present invention, analyze the structure analyzing thing in this sample and illustrate, analyze the wide dynamic range to 5 of thing in this sample to 100,000ng/ml (example 2).That is, conventional analysis bar illustrates following phenomenon, if analyte concentration exceedes certain concentration in sample, then the signal intensity of detection line reduces due to hook effect, as described above.But, the feature of the band for detecting fast of the present invention is, it comprises variable control line further, this variable control line reduces gradually along with the increase of analyte concentration in sample, and illustrate in the dynamic range only with the restriction in the conventional strip of detection line, the scope of hook effect is expanded to, because this variable control line is not by hook effects by this detection line and the combination of this variable control line.But this dynamic range can change according to the type, the amount being fixed on the part on this detecting pad, signal detector etc. of analyzing thing, and therefore dynamic range of the present invention is not limited to above-mentioned scope.
Band for detecting fast of the present invention will be described in more detail.That invents can comprise sample pad for the band detected fast, is incorporated into by the sample comprising target analyte in this sample pad; Pad, one end of this pad is connected to this sample pad; Detecting pad, one end of this detecting pad is connected to the other end of this pad; Absorption layer, one end of this absorption layer is connected to the other end of this detecting pad, provides the driving force that this sample transports from sample pad; With the Solid support on the downside of the band be placed on for detecting fast.Fig. 2 shows the structure of the band for detecting fast of the present invention.
This Solid support can be made up of the material being selected from the colony be made up of nitrocellulose, nylon, PVDF, glass and plastics.Band is attached to increase the permanance of this band on this Solid support, and can processes and store this band simply.In addition, also additional outer box can be installed simply.Wait that the plastic material being used as Solid support can be polypropylene screen, polyester film, polycarbonate membrane, organic film etc., but be not limited thereto.
On the other hand, the invention provides the diagnostic kit comprising the band for detecting fast of the present invention in mounting box, wherein comprise guide and ribbon support part in lower box, and in upper box, form sample reception hole, and result view window is placed on the position corresponding to this detection line and this variable control line.
That invents can be arranged in box for the band detected fast further.For the band being used for detecting fast being placed on correct position and being used for fixing or compressing its multiple guide and/or ribbon support part can be included in this lower box.Optionally, this guide and ribbon support part can also be included in this upper box, are arranged in the position corresponding to this lower box guide and ribbon support part position.That is, this guide and/or ribbon support part can be formed in this lower box, or are formed in this upper box and this lower box, if needed.In addition, can be included in this upper box from this sample reception hole of the signal of this mark and this result view window for detecting in the position corresponding to this detection line, this variable control line and this constant control line.This sample reception hole the form in hole or crack can be formed in one end of this detecting pad, that is, relative with this absorption layer relative to this detection line end and the some place be separated with this detection line fully, can move along this film to make this sample.This result view window can be formed in the some place of this detecting pad placing this detection line and this variable control line, if and/or need, it can be formed to comprise this constant control line with make its be of a size of can with the naked eye or sensor distinguish fully, if further define this constant control line.This detection line, this variable control line and this constant control line can be formed it, as long as can be discernmible under the prerequisite not limiting its size and dimension.
Can utilize typical plastic material, such as polycarbonate, acronitrile-butadiene-styrene (ABS) etc. manufacture this upper box and this lower box, but are not limited thereto.This upper box and this lower box can be manufactured individually, then assembled by typical connecting elements (such as connecting recessed accessory and convex accessory).Alternately, they can manufacture by integral form.
In addition, diagnostic kit can be equipped with standard signal data, and this standard signal packet is containing for containing the detection line of sample of analysis thing and the standard colorimetric table of variable control line that are in multiple concentration.
As used in this, standard signal data refer to the sample relative to the analysis thing comprising variable concentration known, sum up the data of the signal intensity by utilizing this detection line obtained for the band detected fast of the present invention and this variable control line.Can sum up these standard signal data by different way, these data are typically summarized as the color of detection line corresponding to each analyte concentration and variable control line.Similarly, can by carrying out illustration for the litmus paper measuring pH, this litmus paper provides the Standard Colors table of summing up the paper color corresponding to each pH.Specifically, when this employs the standard signal data comprising this Standard Colors table, only with the naked eye can contrast signal intensity and the standard signal data of this analysis thing, it is possible for therefore carrying out simple and rapid semi-quantitative analysis.In this regard, whether this diagnostic kit may be used for utilizing visual check to exist and intensity from the signal of the mark of this detection line and this variable control line, and this makes to perform qualitative or semi-quantitative analysis becomes possibility.In other words, after this sample is applied to this kit, utilize naked eyes whether to be existed by the signal from this detection line and this variable control line and intensity and be equipped with the standard signal data of Standard Colors table to contrast, thus analyze in this sample and analyze thing and whether exist and concentration.
Still in yet another aspect, the invention provides the method utilizing and carry out the qualitative and quantitative analysis analyzing thing in sample for the band detected fast, the method comprises: the pad this sample being incorporated into the prelocalization of this pad or this pad, and allows this sample migration (step 1); Detect and whether exist and intensity (step 2) from the signal of this detection line with the label of this variable control line; And the signal intensity of this detection line and this variable control line and standard signal data are compared the amount (step 3) determining this analysis thing, wherein experience step 1 by each making to comprise in the sample of the analysis thing being in multiple variable concentrations and step 2 obtains this standard signal data.
Also on the other hand, the invention provides in sample, one or more analyze the method for the quantitative of thing or quantitative test, comprise and this sample and the part be adsorbed on Solid support are reacted, it is characterized in that analyzing thing to one is applied with two kinds of different reaction mechanisms.The two kinds of differential responses mechanism being suitable for method of the present invention can be interlayer reaction and competitive reaction.
To carry out the detailed principle of the method for qualitative and/or quantitative test the same with mentioned above to analyzing thing in sample to utilize the band for detecting fast of the present invention.In addition, these standard signal data are the same with mentioned above.
This particular analysis method can be implemented in the following sequence.The first, the fluid sample comprising target analyte can be incorporated in the pad of the prelocalization of this pad or this pad.That is, by this sample is incorporated into this pad, this fluid sample can be incorporated in this band, but can preferably be introduced in the pad of the prelocalization of this pad, such as sample pad.In addition, add damping fluid such as PBS to sample, mix them equably, then this potpourri can be incorporated in this band in the same manner.
When being loaded in (introducing) to the band for detecting fast of the present invention by sample, this sample starts migration, then can be guaranteed the Successful migration of this sample by this constant control line.When ensure that the Successful migration of this sample, check whether the signal of the label from this detection line and this variable control line exists and intensity.This analysis thing of signal designation observed from this detection line is contained in (qualitative analysis) this sample.In addition, can determine to analyze in this sample by the signal intensity and standard signal data contrasting this detection line and this variable control line the concentration (quantitative test) of thing.
In one particular embodiment of the present invention, utilize the band enforcement human immunoglobulin G (IgG for detecting fast further with this variable control line; Analyze thing) analyze.Can confirm that it is possible for the naked eye carrying out quantitative test highly reliably, although under hook effect occurs in the concentration levels of the sample analytes of 5 to 100,000ng/ml scopes (table 1 and Fig. 5).
Sample for immunoassay of the present invention can comprise all biological samples, such as whole blood, haemocyte, serum, blood plasma, marrow, sweat, urine, tears, saliva, skin, mucous membrane, hair etc., above-mentionedly allly all to be separated from mammal, preferably, the mankind.Preferably, this sample is blood.Blood can for by removing serum or blood plasma prepared by haemocyte.If employ whole blood, then can add the composition being used for dissolving haemocyte to damping fluid.These are only for illustrative purposes, but particularly restriction be used for the sample of immunoassay of the present invention.
Immunoassay of the present invention may be used for diagnosing the illness, wherein whole blood is mainly used as sample, and such as malaria antigen (Ag), AIDS virus, the third liver, hepatitis B, syphilis, ulcer cause bacterium, tumor marker (AFP, PSA, CEA), pulmonary tuberculosis, SARS (Severe Acute Respiratory Syndrome), dengue fever, leprosy etc.
Mode of the present invention
Hereafter, the present invention is further described in more detail with reference to example.But these examples are only for illustrative purposes, and the present invention is not intended to limit these examples.
example 1: manufacture the band being used for detecting fast
A. the detecting pad with detection line, variable control line and constant control line is manufactured
Nitrocellulose membrane manufactures three analytical lines.Laminating machine is utilized to carry out layering to this nitrocellulose membrane on plastic clip.After this, utilize automatic distributor respectively using Goat anti human's immunoglobulin like protein of the Ligands as detection line (from Arista company of the U.S., the anti-human IgG of goat), as the human immunoglobulin (IgG) of the 3rd part of variable control line, be assigned on it with the bovine serum albumin(BSA) (BSA) of the reporter molecule as constant control line, then dry 2 days (48 hours) under 25 ~ 30 degrees Celsius.
B. pad is prepared
By pad soak completely in the TRIS buffer (10mM, pH8.5) comprising 0.5%PVA (polyvinyl alcohol (PVA)), then in exsiccator bone dry for the pre-service of this pad.
Prepare the first bond solution by combining the gold grain with about 40nm diameter with Goat anti human's immunoglobulin like protein (the anti-human IgG from goat), and by combination, there is the colloidal gold particles of about 40nm diameter and streptavidin prepares the second bond solution.This first bond solution and this second bond solution are applied to pretreated pad, and in exsiccator bone dry.Subsequently, pad is prepared by being cut into suitable dimension.
C. sample pad is prepared
Sample pad is soaked completely comprise the TritonX-100 of 1%, the NaN3 of 0.5% and 0.1% BSA 0.08M borate buffer in, and in exsiccator bone dry.Subsequently, pad is prepared by being cut into suitable dimension.
D. absorption layer is prepared
Bone dry absorption layer in exsiccator, and use when not processing.Subsequently, pad is prepared by being cut into suitable dimension.
E. for the preparation of the band detected fast
The detecting pad prepared by said procedure, pad, sample pad and the absorption layer structure according to Fig. 4 is assembled.
Namely, the sample pad with sticker is attached to overlapping with one end of pad, one end of detecting pad is attached to overlapping with the other end of this pad, and the other end of this detecting pad is attached to overlapping with one end of the absorption layer with the sticker indicating top.Use cutter this assembly to be cut into the size of about 2 ± 1.0mm, and finally prepare the band in Fig. 4.
In the diagram, the meaning of each reference number is with hereafter identical.
1: sample pad; 16 ± 4 × 4 ± 2mm
2: the pad of anti-human immunoglobulin (Ig) and gold grain; 6 ± 1.0 × 4 ± 2mm
3: nitrocellulose detecting pad; 25 ± 5 × 4 ± 2mm
4: plastic solidification support member
5: absorption layer; 18 ± 4x4 ± 2mm
6: the detection line of anti-human immunoglobulin (Ig)-fixing
7: the variable control line of human immunoglobulin-fixing
8: the constant control line of bovine serum albumin-fixing
example 2: utilize the band being used for detecting fast to carry out immunoassay
Add each 100ml comprised in the sample solution of human immunoglobulin (IgG) and phosphate buffer (PBS) to 96-orifice plate.By utilize 0ng/ml, 5.6ng/ml, 11.2ng/ml, 22.5ng/ml, 45ng/ml, 90ng/ml, 187ng/ml, 375ng/ml, 750ng/ml, 1.5 μ g/ml, 3.12 μ g/ml, 6.25 μ g/ml, 12.5 μ g/ml, 25 μ g/ml, 50 μ g/ml or 100 μ g/ml concentration human immunoglobulin prepare this sample solution.
The band being used for detecting fast of preparation in example 1 is immersed in and comprises in the 96-orifice plate of sample solution, and allow migration 10 minutes.
Fig. 5 is the figure that the development of stratography band is shown according to the concentration as the human immunoglobulin analyzing thing.
In addition, utilize naked eyes detection from the signal intensity of this detection line, this variable control line and this constant control line, and be shown in following table 1.In this regard, by each in color intensity determination signal intensity, wherein the highest color intensity is considered as 5 arbitrarily, and minimum color intensity is considered as 0.
Table 1
[table 1]
| The concentration (ng/ml) of target | Constant control line | Variable control line | Detection line |
| 0 | 3 | 5 | 0 |
| 5.6 | 3 | 5 | 0.5 |
| 11.2 | 3 | 5 | 1 |
| 22.5 | 3 | 5 | 2 |
| 45 | 3 | 5 | 2 |
| 90 | 3 | 5 | 2 |
| 187 | 3 | 4 | 3 |
| 375 | 3 | 4 | 4 |
| 750 | 3 | 4 | 4 |
| 1500 | 3 | 3 | 3 |
| 3120 | 3 | 3 | 3 |
| 6250 | 3 | 2 | 2 |
| 12500 | 3 | 1 | 2 |
| 25000 | 3 | 0.5 | 2 |
| 50000 | 3 | 0 | 2 |
| 100000 | 3 | 0 | 1 |
As shown in table 1, when the concentration analyzing thing (IgG) in sample increases, the signal intensity of this detection line increases gradually, and observes peak signal when about 375ng/ml to 750ng/ml.Afterwards, signal intensity reduces gradually, and this gives the credit to hook effect mentioned above.Therefore, thing is not analyzed by only utilizing the signal intensity of this detection line to carry out quantitative test.
Such as, if the signal intensity of this detection line is 2, then analyze the concentration of thing (IgG) in sample as seen near 40ng/ml and 10 μ g/ml.Although this detection line shows identical signal intensity, this analyte concentration can be different.
But when the concentration analyzing thing (IgG) in this sample increases, the signal intensity of this variable control line reduces gradually.In details, visible, under analyte concentration is 0 to 90ng/ml situation, signal intensity almost remains on highest level 5.So, if analyte concentration is low, in detection line, hook effect is not had to occur, and therefore can by the signal intensity determination analyte concentration of this detection line.If this analyte concentration increases, then the signal intensity of this detection line becomes irregular due to hook effect.But the signal intensity of this variable control line reduces continuously, therefore, this variable control line determination analyte concentration can be passed through.
Such as, if the signal intensity of this detection line is 2, then this analysis thing can have two concentration of about 40ng/ml and 10 μ g/ml, as described above.But this variable control line shows the signal intensity of 5 or 2 relative to these two concentration, indicates the quantitative test of this concentration to be possible, is different from the conventional method being only suitable for detection line.
In addition, can find, the band for detecting fast of the present invention has 5ng/ml to 100, the wide dynamic range of the analysis thing of 000ng/ml.
Industrial applicibility
Because the band for detecting fast of the present invention has variable control line, so to analyze the difficulty of thing in quantitative test and utilize naked eyes to carry out quantitative test highly reliably together with qualitative analysis in simple and quick mode be possible because hook effect overcomes.In addition, there is effect as follows: the dynamic range analyzing thing can be improved by the complementary effect of detection line and variable control line.
Claims (18)
1. one kind has the band for detecting fast of pad and detecting pad, wherein said pad has and marks with the first part of the first scheduled volume of analyte response and the input that is attached to described first part, described first part and described be marked at sample migration before or link each other when sample migration to form bond, and described bond can move to detecting pad when described sample migration
Wherein said detecting pad has detection line separated from one another and variable control line,
Wherein said detection line is fixed together with the Ligands with analyte response in described sample, and
Wherein said variable control line is fixed together with the 3rd part of the second scheduled volume, described 3rd part is equal to and the described analysis thing of described first ligand reaction or its analog, thus can react with not being attached in described sample the naked bond analyzing thing.
2. the band for detecting fast according to claim 1, analysis thing in a part for first part of wherein said first scheduled volume and described sample reacts, with thus formed compound, wherein said analysis thing is attached to the bond with described first part and described label, and the amount of the compound formed increases along with the increase of analyte concentration in described sample, but the amount of described naked bond reduces.
3. the band for detecting fast according to claim 2, wherein there is described first part, the described compound of described label and described analysis thing reacts with the described Ligands of catching at described detection line place, be not attached to the analysis thing in described sample and the described naked bond with described first part and described label reacts with described 3rd part of catching at described variable control line place, and can determine whether the analysis thing in described sample exists or its amount by the signal measured from described detection line and described variable control line, or both above-mentioned.
4. the band for detecting fast according to claim 3, the signal intensity of wherein said variable control line reduces along with the increase of analyte concentration in described sample.
5. the band for detecting fast according to claim 4, signal intensity wherein from described variable control line remains constant level, until the concentration analyzing thing in described sample reaches M value, but when described concentration reduces gradually more than described signal intensity during M value, and finally converge to 0, wherein when all 3rd parts fixed with described second scheduled volume are reached capacity by the reaction of described naked bond, M value is analyze the Cmax of thing in described sample.
6. the band for detecting fast according to claim 1, wherein said detecting pad has the constant control line for determining described sample migration further, and described constant control line is fixed together with reporter molecule.
7. the band for detecting fast according to claim 1, comprising:
Sample pad, is incorporated in described sample pad by the sample comprising analysis thing to be analyzed;
Pad, one end of this pad is connected to this sample pad;
Detecting pad, one end of this detecting pad is connected to the other end of this pad;
Absorption layer, one end of described absorption layer is connected to the other end of described detecting pad, provides the driving force that described sample transports from described sample pad; With
Be placed on the Solid support on the downside of the band for detecting fast.
8. the band for detecting fast according to claim 7, wherein said support member is made up of the material being selected from lower group, and this group is made up of the following: nitrocellulose, nylon, Kynoar, glass and plastics.
9. the band for detecting fast according to claim 1 is wherein the first part, described Ligands and described 3rd part are protein, antigen, antibody, DNA, RNA, PNA or fit.
10. the band for detecting fast according to claim 1, wherein said label is aurosol, emulsion particle, colored ps particle, enzyme, fluorescent pigment, conducting polymer, luminescent substance or magnetic-particle.
11. bands for detecting fast according to claim 1, described band is immunochromatography bar.
12. 1 kinds of diagnostic kits of band for detecting fast comprised according to any one of claim 1 to 11, described diagnostic kit is additionally arranged in box, wherein said kit has guide and ribbon support part in lower box, there is sample reception hole in upper box, and result view window is placed on the position corresponding to described detection line and described variable control line.
13. diagnostic kits according to claim 12, wherein said kit is equipped with standard signal data, and described standard signal packet is containing for containing the described detection line of sample of described analysis thing and the standard colorimetric table of described variable control line that are in multiple concentration.
Whether 14. diagnostic kits according to claim 13, wherein can be existed and intensity by the signal utilizing described naked eyes to carry out the described mark that qualitative or semi-quantitative analysis checks from described detection line and described variable control line.
15. 1 kinds utilize the method for carrying out the qualitative or quantitative test analyzing thing for the band detected fast according to any one of claim 1 to 11, said method comprising the steps of:
This sample is incorporated into this pad or in the pad of the prelocalization of this pad, and allows this sample migration (step 1);
Whether the signal detected from the mark of described detection line and described variable control line exists and intensity (step 2); With
By described detection line be the amount that signal intensity and the standard signal data of variable control line compare to determine described analysis thing, wherein experience step 1 by each making to comprise in the described sample of the described analysis thing being in multiple variable concentrations and step 2 obtains described standard signal data (step 3).
Whether 16. methods according to claim 15, wherein in step 2, utilize opacimeter inspection to exist and signal intensity from the signal of the label of described detection line and described variable control line.
17. 1 kinds in qualitative or quantitative test sample, one or more analyze the methods of things, described method comprises makes described sample and the part be adsorbed on Solid support react, and it is characterized in that analyzing thing to one is applied with two kinds of different reaction mechanisms.
18. methods according to claim 17, wherein said two kinds of different reaction mechanisms are interlayer reaction and competitive reaction.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR10-2013-0064207 | 2013-06-04 | ||
| KR20130064207 | 2013-06-04 | ||
| PCT/KR2014/004961 WO2014196803A1 (en) | 2013-06-04 | 2014-06-03 | A strip for rapid testing having a variable control line and a diagnosis kit using the same |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105229468A true CN105229468A (en) | 2016-01-06 |
Family
ID=52008377
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201480020367.8A Pending CN105229468A (en) | 2013-06-04 | 2014-06-03 | Have variable control line for the band that detects fast and the diagnostic kit using this band |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP3004879A4 (en) |
| JP (1) | JP2016520200A (en) |
| KR (1) | KR101540608B1 (en) |
| CN (1) | CN105229468A (en) |
| WO (1) | WO2014196803A1 (en) |
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Also Published As
| Publication number | Publication date |
|---|---|
| EP3004879A4 (en) | 2017-01-11 |
| JP2016520200A (en) | 2016-07-11 |
| KR20140143120A (en) | 2014-12-15 |
| KR101540608B1 (en) | 2015-08-03 |
| WO2014196803A1 (en) | 2014-12-11 |
| EP3004879A1 (en) | 2016-04-13 |
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