CN101021531B - Detecting device containing non-protein trapping matter on control region - Google Patents
Detecting device containing non-protein trapping matter on control region Download PDFInfo
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- CN101021531B CN101021531B CN2006100503082A CN200610050308A CN101021531B CN 101021531 B CN101021531 B CN 101021531B CN 2006100503082 A CN2006100503082 A CN 2006100503082A CN 200610050308 A CN200610050308 A CN 200610050308A CN 101021531 B CN101021531 B CN 101021531B
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Abstract
The invention provides a detection device to detect if the sample has the analyte. The device includes the regent bar which supports the liquid sample flow; the bar includes the detection region which includes a special combinative molecule and the control region which includes one or more non-protein catch; in some material operation, one or more non-protein flocculating regent or the coagulation are on the control region or the masculine control region. The production cost is low but not change the detecting result.
Description
Technical field
The invention belongs to the diagnostics classes device, specifically, is about a kind of novel diagnostic reagent bar, comprises that on the control area of reagent strip a certain amount of one or more non-albumen catch material.
Background of invention
Utilize the dry chemistry test paper slip of immunology principle reaction to be used in a large number in the clinical diagnosis, for example the very early pregnancy reagent strip detects HCG.This type test strips and being used in of they all have description in a lot of patent documentations; For example China has been authorized to utility model patent 01239923.X and 02202021.7; And application publication 02122907.4 and 02139704.X, 01131834.1 or the like that kind that disclosed.For example; The most common test strips of utilizing the single stage method detection of sandwich principle; On the sample region of acceptance on the test strip, add the sample that institute will detect, because capillary layer suction effect is vertically flowed forward along test paper, pass through marked region in this sample; If there is analyte in the sample, another material specific bond forms compound on this analyte and the marked region; This compound is along with liquid continues to flow forward, and through including the surveyed area of catching the analyte zone, again with the catches specific bond of catching on the analyte zone, thereby compound is hunted down, and makes at surveyed area to accumulate.Another material-specific on the marked region can be labeled the material mark; This mark substance can be the band colored particle of the known non-water-soluble of prior art; For example gold grain colloid and latex particle; Also can be fluorescence labeling, can also be the marking particle of water-soluble, marking particle that is for example formed by the dextran polymerization or the like.On marked region, contain the band colored particle, a colored symbol then on surveyed area, occurs, this symbol both can directly with the naked eye just can be read the result, also can by instrument qualitative and quantitative more accurately read the result.This analyte and the pairing between the catches specific bond of catching on the analyte zone; And analyte and the combination that has between the material of mark are the known materials of prior art; These both combine can the direct specific bond between them also can be indirect specific bond; Common like double-antibody sandwich, double antigens sandwich or its indirect method also have competition law or the like.These pairings have a lot of modes, and for example antigen matches with its antibody, antibody and antiantibody pairing; Antibody and haptens pairing; Pairing between the antibody of biotin and antibiotin, biotin and antibody pairing, and a plurality of combinations of pairs of formation between them or the like; Antibody also comprises the antibody of antibody fragment, the antibody in for example anti-Fc site or the like.
Simultaneously; The downstream of catching the analyte zone on these reagent strips also have a testing result control area; On this zone, also be fixed with a certain amount of antibody or special protein substance, these antibody or special protein substance (albumin A or protein B) can be caught labeled complex matter unnecessary in the solution and accumulation in the testing result control area.Generally, color in the testing result control area, occurs and represent that then testing result on the surveyed area is effectively, otherwise be invalid, for example U.S. Pat 5,541,059 that kind that disclosed with European patent EP 0 560 411.In addition, in some patent documentations, disclose and be fixed on antibody on the positive control area, no matter whether have analyte in the sample; The color lines on positive control area, all occur, these lines and specific bond analyte zone intersect to form special symbol that can Direct Recognition, and for example U.S. Pat 4; 916,056, US 5,160; 701 that kind that disclosed with US 5,075,078.From prior art; Fixing special antibody or other similar differential protein material are as catching material by the method for generally using on testing result control area or positive control area; Therefore; In producing in a large number, on the control area, can improve the cost of producing product greatly on the one hand because of the specific antibody of use high price or albuminoid material; Also can cause the waste of the specific bond molecule that has mark substance in a large number on the other hand, because will form the control lines, the general specific bond molecule that has mark substance on reagent areas is excessive.
Openly apply for a patent in the U.S. and to have disclosed another kind of testing result control area in 2004/0161859; Before reagent strip is not used; On the testing result control area on the reagent strip, just there have been blue lines; When reaction was carried out, these blueness lines will become redness and represent that reaction has finished or testing result is effective.Though this reagent strip has reduced production cost,, just there have been tangible lines in this reagent strip on the testing result control area before not using, be easy to like this let the user produce illusion, thinks that this reagent strip was used.
At United States Patent (USP) 6,855,561 have disclosed and have utilized the pick-up unit of a kind of fixing of dye on positive control area.In embodiment, this device is not participated in before the reaction, and this dyestuff that itself has a kind of color is hidden in below the nitrocellulose membrane; When detecting, become transparent owing to nitrocellulose filter is wet by water, thereby the dyestuff that is hidden under the film is displayed and combine the certain discernible shape of zone composition of analyte.In actual production, use special devices and need complicated loaded down with trivial details technology just can produce the actual product that business demand such as this patent disclose that satisfies.
Apply for a patent in China; Application number be disclosed in 200510049177.1 comprising when one or more are being done it being that a kind of color is the material of another kind of color on the positive control area in wet, this positive control area and analyte calmodulin binding domain CaM can form certain discernible symbol.In actual production; To show that under dried and wet two kinds of different situations direct processing of material of different colours is the difficult thing of comparison on nitrocellulose filter this; Be that this material can freely spread on film the efficient of processing is reduced greatly because directly handle the solution that contains this material on film.
In order to overcome these defectives of the prior art and deficiency, processing flocculation reagent or coagulum can well reach the coml requirement on our control area of very surprised discovery in this type pick-up unit.This type flocculation reagent or coagulum are the reagent that wastewater treatment is often used, their low prices, and obtain easily.So both can practice thrift in a large number because of buying the expensive cost that antibody or albumen spent; On the other hand; In a concrete embodiment, do not need special devices and complicated technology just can directly be fixed on the positive control area of formation on the nitrocellulose filter to this flocculate reagent or coagulum.
Technical scheme
The present invention provides the pick-up unit that whether contains analyte in a kind of test sample, and pick-up unit comprises the reagent strip of supporting that fluid sample flows; Comprise surveyed area and control area on this reagent strip; Comprise a kind of molecule of specific bond at surveyed area, comprise the catches of one or more non-protein ingredients in the control area.
In one embodiment; On the water absorptivity reagent strip, have only surveyed area and testing result control area; On surveyed area, comprise a specific bond molecule, on the testing result control area, comprise the flocculation reagent or the coagulum of one or more non-protein ingredients.When detecting; Can be earlier the molecular mixing of another specific bond of sample that will detect and tape label material; If contain the material that will detect in the sample; Change color then on surveyed area, can occur and show whether there is analyte in the sample, change color on the testing result control area, occur and show whether this testing result is effective.Usually, there on the testing result control area, have color this testing result of expression to occur to be effective, otherwise be invalid testing result.
In another embodiment; Support that the reagent strip of liquid flow is the cellulose nitrate reagent strip; On surveyed area, be fixed with the molecule of specific bond analyte; Accomplish the molecule that the necessary reagent areas of reaction comprises another specific bond analyte that has mark substance, on the testing result control area, be fixed with a certain amount of non-albumen flocculation reagent or coagulum.
In another embodiment, also comprise reagent areas on the reagent strip, on reagent areas, comprise two zones, one is the zone that has dyestuff, and another zone is a marked region, on marked region, comprises another specific bond molecule that has mark substance; The molecule that on surveyed area, comprises a specific bond comprises a certain amount of one or more non-albumen flocculation reagent or coagulums in the testing result control area.In a concrete embodiment; The specific bond molecule that has mark substance can be antibody, antibody fragment; One of biotin or nucleic acid (DNA, RNA) fragment, the specific bond molecule on the surveyed area can be one of antibody of another antibody, antibody fragment, anti-biotin antibodies or anti-nucleic acid.
In another embodiment; This reagent strip is made up of water-absorbing material, and it comprises the sample region of acceptance, accomplishes the necessary reagent areas of reaction, surveyed area, testing result control area and suction zone; On the testing result control area, contain a certain amount of non-albumen flocculation reagent or coagulum; The molecule that on surveyed area, contains specific bond comprises marked region and dyestuff zone on reagent areas, on marked region, contain another molecule that has the mark substance specific bond.
In another embodiment; Reagent strip is the nitrocellulose filter reagent strip; Reagent areas comprises specific bond molecule and a certain amount of dyestuff that has mark substance; On surveyed area, comprise specific bond analyte zone and positive control area, on positive control area, comprise a certain amount of non-albumen flocculation reagent or coagulum, the zone of specific bond analyte and positive control area intersect to form certain shape each other.In this embodiment, can also comprise testing result control area and suction zone in the downstream of surveyed area, on the testing result control area, comprise a certain amount of non-albumen flocculation reagent or coagulum.Simultaneously; In another embodiment; Can fix some protide catches on the positive control area; Antibody of antibody, antibody fragment, albumin A, Protein G or anti-albumin A or the like for example, the flocculation reagent or the coagulum of more fixing non-protein ingredients on the testing result control area, the zone of specific bond analyte and positive control area intersect to form each other+/-, the shape of identification easily such as Y, X.In this embodiment in another practical implementation example; Reagent strip comprises that being from upstream to downstream along liquid flow direction is followed successively by nitrocellulose filter and the adsorptive pads that sample is accepted pad, label pad, contained surveyed area and testing result control area, and these parts link to each other and can let liquid flow to the other end from an end through the capillary absorption affinity; On sample is accepted to fill up, handling has some reagent to come the pH value of conditioned reaction; On label pad, handle the specific bond molecule that has mark substance is arranged; On nitrocellulose filter, handle molecule and the positive control area that the specific bond analyte is arranged; On positive control area, handling has a certain amount of non-albumen flocculation reagent or coagulum, and the zone of positive control area and specific bond analyte intersects to form certain shape each other.These non-albumen flocculation reagent or coagulum can be some inorganic polymer flocculation reagent or organic polymer flocculation reagent.Mark substance on the reaction reagent zone can be colored colloidal solid, like gold grain colloid, latex particle; Can also be to apply for a patent the 99809129.4 water-soluble mark substances that disclose like China invention.
On the other hand, the invention provides the method that detects analyte in the sample.In one embodiment; To pick-up unit dropping liquid sample body; Make sample pass through marked region, surveyed area and testing result control area in order from the sample region of acceptance of reagent strip; When flow of liquid is crossed the testing result control area; Be dissolved in the catches that the specific bond molecule is caught on the part zone not to be detected in the liquid mark substance is fixed on one or more the non-protein ingredients on the testing result control area and catch, change color on the testing result control area, occurs and represent that this testing result is effective.In another concrete embodiment, non-protein molecular is flocculation reagent or the coagulum that organic or inorganic is dissolved in water.
In another embodiment, to detectable bar sample region of acceptance dropping liquid sample body, this liquid sample relies on the capillary absorption affinity to flow through marked region, dyestuff zone, surveyed area and testing result control area successively.When liquid passed through on the dyestuff zone, this dyestuff can and flow to the testing result control area by the liquid sample dissolving, and this dyestuff is caught by the coagulum of non-protein ingredient or flocculation reagent in the testing result control area.In another concrete embodiment; Surveyed area comprises the zone and the positive control area of specific bond analyte; Positive control area and specific bond analyte zone intersect to form certain shape each other, on positive control area, handle flocculation reagent or the coagulum that a certain amount of non-protein ingredient is arranged.When the flow of liquid that is dissolved with dyestuff was crossed positive control area, flocculation reagent on positive control area or coagulum can be caught the dyestuff in the liquid and demonstrated change color.If there is not analyte in the sample, color lines negative signs (-) just only appear on surveyed area, and the expression testing result is negative; If there is analyte, on specific bond analyte zone the color lines appear then, and the lines on these lines and the positive control area intersect to form positive sign (+), the positive result of ecbatic.The flocculation reagent of the non-protein ingredient on the testing result control area or coagulum also can catch in the liquid dyestuff and on the testing result control area Show Color, represent that this testing result is effective result, otherwise this testing result is invalid.
In another embodiment; On the water absorptivity reagent strip, have only surveyed area and testing result control area and sample region of acceptance; On surveyed area, comprise a specific bond molecule, on the testing result control area, comprise the flocculation reagent or the coagulum of one or more non-protein ingredients.When detecting; Can be earlier the molecular mixing of another specific bond of sample that will detect and tape label material; Be inserted into the sample region of acceptance of reagent strip in the liquid sample then and go; Want analyte if contain in the sample, change color then on surveyed area, can occur and show whether there is analyte in the sample, change color on the testing result control area, occurs and show whether this testing result is effective.Usually, there on the testing result control area, have color this testing result of expression to occur to be effective, otherwise be invalid testing result.
Description of drawings
Fig. 1 is for the invention provides an embodiment.On reagent strip 1, comprise sample region of acceptance 25, reagent areas 115, surveyed area 95, testing result control area 85 and suction zone 65; Arrow express liquid flow direction includes a certain amount of non-protein ingredient coagulum or flocculation reagent on testing result control area 85.
Fig. 2 is for the invention provides another embodiment.Compared to Figure 1, difference is, on the reagent areas on the reagent strip 2 115, comprises the marked region 15 of being with colored particle and the dyestuff zone 35 that has dyestuff, on testing result control area 85, comprises the flocculation reagent or the coagulum of non-protein ingredient.
Fig. 3 is for the invention provides another embodiment.Compare with Fig. 2; On surveyed area 95, comprise a positive control area 55 and analyte calmodulin binding domain CaM 45; Testing result control area 85; Wherein on testing result control area 85 and positive control area 55, all handled a certain amount of coagulum or flocculation reagent, simultaneously positive control area 55 intersects to form one ten sub-frame shape (+) with the analyte calmodulin binding domain CaM.
Fig. 4 the invention provides an embodiment shown in Figure 3.Reagent strip 4 is linked to each other by sample blank film 425, marker slip 4115, nitrocellulose filter 475 and suction sheet 465; Liquid can flow on the absorbing sheet 465 from sample blank film 425, and wherein surveyed area 495 is positioned on the nitrocellulose filter 475 with testing result control area 485.Handling on the marker slip 415 has specific bond molecule and the dyestuff that has the color mark material, at the molecule that combines to comprise on the analyte zone 445 the specific bond analyte.On positive control area 455 and testing result control area 485, handle a certain amount of coagulum or flocculation reagent are arranged.
Fig. 5 is for the invention provides another embodiment; Compare with Fig. 3; On reagent areas 115, include only a marked region 15, do not have testing result control area 85 and dyestuff zone 35 simultaneously, on positive control area 55, comprise a certain amount of coagulum or flocculation reagent.
Fig. 6 the invention provides an embodiment shown in Figure 5; Compare with Fig. 4; There has not been testing result control area 485 on this reagent strip 6; On marker slip 4115, include only the specific bond molecule of band colored particle, on positive control area 455, comprise a certain amount of non-albumen coagulum or flocculation reagent.
Fig. 7 the invention provides a concrete embodiment, and the embodiment difference of showing with Fig. 2 is that handling on the marked region 15 has the specific bond molecule that has the color water-soluble dye.
Fig. 8 is for the invention provides another embodiment, and the embodiment difference of showing with Fig. 7 is do not have the testing result control area to exist on the reagent strip 8, and on positive control area, handling has a certain amount of coagulum or flocculation reagent.
Description of reference numerals
Reagent strip 1,2,3,4,5,6,7,8, sample region of acceptance 25, reagent areas 115, marked region 15; Dyestuff zone 35, surveyed area 95, analyte calmodulin binding domain CaM 45, testing result control area 85; Positive control area 55, suction zone 65, sample blank film 425; Marker slip 4115, nitrocellulose filter 475, the surveyed area 495 on the nitrocellulose filter; Testing result control area 485 on the nitrocellulose filter, the analyte calmodulin binding domain CaM 445 on the nitrocellulose filter, the positive control area 455 on the nitrocellulose filter.
Embodiment
In the detailed description below, the subsidiary reference word of legend is a part here, and it is explained to illustrate the mode that the present invention can actable specific concrete scheme.We do not get rid of the present invention and can also carry out other concrete scheme and under the situation of claim scope of the present invention, change structure of the present invention.
Surveyed area
Here said surveyed area is meant on this zone, can react to detect whether there is analyte of interest matter in the sample.Here said reaction can be immune response, like single stage method immunoreagent bar or the like.Like Fig. 1, Fig. 2 and shown in Figure 7, contain a surveyed area 95 on the reagent strip, on surveyed area 95, fixed some specific bond molecules, on this zone, detect whether contain analyte in the sample.
In one embodiment, on surveyed area 95, fix the molecule of a specific bond analyte, comprise another molecule of the specific bond analyte that has mark substance on the reagent areas 115.When containing analyte in the sample; Another specific bond molecule forming composite matter of a part on analyte and the reagent areas; This compound is through on the surveyed area; Be fixed on another specific bond molecule trapping on the surveyed area and accumulate on the surveyed area,, show whether analyte is present in the sample so change color on surveyed area, also can occur owing to have color on the compound.In embodiment, another specific bond molecule of tape label material can be antibody or antibody fragment, and another specific bond molecule that is fixed on the surveyed area also can be antibody or antibody fragment.Be generally double antibody sandwich method or dual-antigen sandwich method detects.
In another embodiment; Another molecule that does not contain the specific bond analyte that has mark substance on the reagent strip; But before reaction; Earlier form WS to sample and another molecular mixing that has mark substance, and then be inserted into reagent strip and go in this mixed solution perhaps to be added drop-wise to mixed solution on the reagent strip, change color also can occur at the surveyed area of reagent strip like this and show whether there is analyte in the sample.
In another embodiment, the specific bond molecule that has mark is the analog of analyte, on surveyed area, is fixed with the molecule of specific bond analyte.When containing analyte in the sample; The similar substance of analyte and tape label can be vied each other and combined the specific bond molecule on the surveyed area; Like this on surveyed area can because analyte what and different depth colors occur and show what of analyte; If analyte is many more, the color relation that on surveyed area, occurs is shallow more, otherwise just dark more.The for example common competition law that utilizes carries out drugs detectable bar.
Shown in Fig. 3-6,8.In other embodiments; This surveyed area 95 comprises positive control area 55 and analyte calmodulin binding domain CaM 45; On positive control area, can be fixed with protein ingredients such as antibody, antibody fragment and catch material; Also can fix the material of catching of some non-protein ingredients, for example inorganic coagulum or flocculation reagent catches comprise the molecule of specific bond analyte on the analyte calmodulin binding domain CaM.But the certain distinguished symbol of this positive control area and the interlaced formation of analyte calmodulin binding domain CaM comes directly to show the result who detects.For example form positive sign (+) or negative sign (-) or the like (Fig. 5 and Fig. 3).In this embodiment, no matter combine the analyte zone whether to have color to occur, all there is color to occur on this positive control area.Specifically, if there is analyte in the sample, the symbol (+) of positive findings just appears on surveyed area 95 being expressed as, if do not have analyte then the symbol (-) of negative findings occurs representing.
The technology of fixing a certain amount of specific bond molecule is that persons skilled in the art can be known with utilize this technology to detect whether having analyte in the sample on the surveyed area; Explanation and elaboration that these technology are all detailed in the prior art; For example U.S. Patent number 5,075, and 078; 5,541,059 that kind that disclosed with European patent EP 0 560 411 grades.
The control area
Here there is the effect of several aspects said control area.Whether the one, whether can be used as the testing result control area, to control the testing result that is presented on the surveyed area effective, or be presented at the process of reacting on the surveyed area and how long accomplish or accomplished.Can also be as the positive findings control area; Another be exactly with surveyed area on another analyte calmodulin binding domain CaM form certain shape and come reaction detection result directly perceived; When containing analyte in the sample; The positive findings that shows just directly perceived, when not having analyte in the sample, the negative findings that shows just directly perceived.Give explanation below in detail.
The testing result control area.In the prior art, the antibody of some protein ingredients or antibody fragment are fixed on the testing result control area in surveyed area downstream, and the excessive specific bond molecule that has mark substance is processed at the upstream region of surveyed area.When detecting; These that do not have that another specific bond molecule on the zone to be detected caught have specific bond molecules of mark substance these protein ingredients on can control area as a result to be detected catches material and catches, thereby shows that this testing result is effective.On the contrary, if on the testing result control area, do not have change color, represent that this testing result is invalid, this detection must be from new detection so.On the other hand; The indicative function of reaction process on the surveyed area can also be served as in the testing result control area; For example on the testing result control area, change color occurred, can think that the reaction on the surveyed area is through with, result displayed is the net result that detects on the surveyed area.The distance of testing result control area and surveyed area is also can regulate arbitrarily according to demands of different; For example the testing result control area is can the distance detecting zone distant, and approximately flowing to the testing result control area apart from liquid from surveyed area needs 5 minutes to 10 minutes distance.The setting of this distance can be according to freely designing in the cellulosic speed of flow of liquid pernitric acid.Like this, when change color appearred in the testing result control area, the reaction of carrying out on the expression surveyed area had continued about 5-10 minute.Control area on the reagent strip can be one or more; Process for control detection reaction more accurately; Can locate to set up first testing result control area in about 5 minutes in downstream, distance detecting zone; Locating to set up second testing result control area in 10 minutes in distance, it is about 5 minutes to represent that when color appears in first testing result control area reaction on the surveyed area has continued, and representes that when change color has appearred in second surveyed area 2 reaction on the surveyed area has continued 10 minutes; Think that perhaps the result on the surveyed area is effectively in 10 minutes, it is invalid having crossed 10 minutes results on the expression surveyed area.
The present invention said " upper reaches " or " downstream " are meant the upper reaches or downstream on the liquid flow direction.Under the normal condition, liquid flow to downstream area from upstream region.For example liquid begins to flow from the sample region of acceptance; Pass through surveyed area and testing result control area flow further downstream successively; Then the sample region of acceptance is positioned at the upper reaches of surveyed area; Surveyed area is positioned at the upper reaches of testing result control area, and the testing result control area is positioned at the downstream of sample region of acceptance and surveyed area.
Positive control area.Simultaneously, in the prior art, the antibody of a large amount of protein ingredients or antibody fragment are fixed on the positive control area on the surveyed area, regardless of testing result, on positive control area, all change color can occur.Another analyte calmodulin binding domain CaM on the surveyed area and positive control area are formed certain shape and are come reaction detection result directly perceived.These antibody or antibody fragments that are fixed on the positive control area also are to be used for catching the superfluous specific bond molecule that has mark.Like this, in a large amount of production, need the antibody or the antibody fragment of a large amount of protein ingredients, simultaneously, also need a large amount of specific bond molecules that has the color mark material.The cost of the reagent strip product of producing like this can significantly improve, because production or purchase protein antibodies or antibody fragment are very high with the cost that has the specific bond molecule of mark substance.In addition; In the prior art some chemical substance treatment on positive control area; For example United States Patent (USP) 6; 855,561 disclose in 2006/0029924 with U.S. Patent application that being utilized in of describing handled some chemical dyes on the positive control area or some utilize chemical reaction to come Show Color to change, thereby make up visual display detecting result with the analyte calmodulin binding domain CaM.But in real process; Directly handling these chemical substances is very difficult on nitrocellulose filter; Handle superincumbent chemical substance and spread easily, because chemical reaction also can influence testing result, the cost of producing this product in addition is also very high simultaneously.
Utilize reagent strip provided by the invention, on the control area, handle certain non-albumen and catch material and can make the reagent strip of production with low cost, produce and can not influence the quality of reagent strip easily.In the embodiment as shown in Figure 1, on reagent strip 1, comprise a testing result control area 85, on the testing result control area, be fixed with the coagulum or the flocculation reagent catches of a certain amount of non-protein ingredient.On reagent areas 115, handle the specific bond molecule that movably has mark substance is arranged.When these specific bond molecules that have mark substance are taken to 85 places, testing result control area by liquid, be fixed on the molecule that the coagulum on the testing result control area or the reagent that flocculates are caught these tape label materials.Change color so just on the testing result control area, occurs and show that testing result is effectively, otherwise testing result is invalid.In another concrete embodiment; Have the coagulum of a certain amount of non-protein ingredient or the testing result control area of flocculation reagent and be positioned at the about 10 minutes distance in surveyed area downstream, when on the testing result control area, change color occurring, then represent to react and carried out 10 minutes.
In another embodiment as shown in Figure 2; On testing result control area 85, be fixed with a certain amount of coagulum or flocculation reagent; On the dyestuff zone 35 at surveyed area 95 upper reaches, handle a certain amount of dyestuff is arranged; Do not have any specific bond molecule on this dyestuff, on the marked region 15 at the upper reaches, dyestuff zone, handle the specific bond molecule that has colored particle simultaneously.When detecting; Dyestuff is taken on the testing result control area 85 by coagulum or flocculation reagent place by liquid and catches and accumulate; Thereby change color on the testing result control area, occurred and represented that testing result is effectively, if do not have change color then represent that testing result is invalid.Simultaneously, specific bond molecule on the surveyed area 95 and the molecule that has mark substance on the marked region 15 carry out immune response and detect whether there is analyte in the sample.In an embodiment, the mark substance on the marked region is granular metallic colloid or latex colloid, and the specific bond molecule on the mark substance can be antibody or antibody fragment, also can be antigen or antigen fragment or the like.In another embodiment, the embodiment difference of showing with Fig. 2 is that handling on the marked region 15 has the specific bond molecule (Fig. 7) that has water-soluble dye.
In another embodiment as shown in the figure; The zone 45 and positive control area 55 of on surveyed area 95, containing the specific bond analyte; One testing result control area 85 is arranged in the downstream of surveyed area 95; At the surveyed area upper reaches reagent areas 115 is arranged, comprise marked region 15 and dyestuff zone 35 on the reagent areas.Wherein, on positive control area 55 and testing result control area 85, be fixed with coagulum or flocculation reagent catches respectively, the zone 45 and the positive control area 55 of specific bond analyte intersect to form ten sub-frame shapes each other.When detecting, the dyestuff on the dyestuff zone is caught the positive control of formation lines by the coagulum of positive control area or the reagent that flocculates, and coagulum on the control area as a result 85 to be detected or flocculation reagent are caught the formation result and controlled lines.Simultaneously, the specific bond molecule that has the color mark material on the marked region is just participated in and the reaction of specific bond analyte comes testing result, participates in positive control line of formation or testing result control line and need not handle excessive mark substance.The binding molecule that has mark substance that a part is expensive has so just been practiced thrift by good.
In another concrete embodiment; Also can handle the antibody or the antibody fragment of some protein ingredients on the positive control area 55; The albumen that perhaps other can the capture of labels material is caught material, on the testing result control area, is fixed with coagulum or flocculation reagent simultaneously.
In another embodiment shown in Figure 5, on surveyed area, comprise the zone of containing specific bond analyte molecule and with positive findings control area 55, on positive structure control zone, handling has a certain amount of coagulum or flocculation reagent to catch.Simultaneously, a processing has the transportable specific bond molecule that has the color mark material on reagent areas.Like this, when detecting, coagulum on the positive control area or flocculation reagent are caught unnecessary mark substance and are formed the control lines.These positive control lines can show testing result with the symbol that the zone formation of specific bond analyte can be discerned.
Non-albumen catches
The catches that material replaces to non-protein ingredient of catching of the antibody on the prior art control area or antibody fragment or some protein ingredients, greatly reduce the cost of reagent strip like this and don't change sensitivity and the specificity that reagent strip detects.The a large amount of utilizations of pollutant quilt in field of waste water treatment of adopting flocculation reagent or coagulum to separate and removing the colloidal state in the waste water.The present invention utilize flocculation reagent or coagulum to have to condense with the precipitated liquid sample body in this character of particle form change color in the control area.Absorption that this flocculation reagent or coagulum cause and deposition are nonspecific absorption and deposition.Here non-specific is that the particulate material that has in the solution can and absorb by flocculation reagent or coagulum deposition.Here said non-albumen is and the prior art relative material of antibody, antibody fragment or albumin A, protein B commonly used; Be not meant that these flocculation reagent or coagulum do not contain any protein substance; But these flocculation reagent or coagulum substantially are not to catch other specific bond molecules in the solution with the specific bond mode, but they come the particle or the particle matter in the solution of catching of non-selectivity with the mode of non-specific bond.
In a concrete embodiment, as shown in Figure 1, fixing a certain amount of flocculation reagent or coagulum on testing result control area 85, the special molecular of a certain amount of band color colloidal solid of 115 processing on the reagent areas at surveyed area 95 upper reaches.When detecting, flow through reagent areas 115 successively from the liquid of sample region of acceptance 25, surveyed area 95 and testing result control area 85.When flowing through testing result control area 85, be comprised in flocculation reagent or the coagulum that the band color colloidal solid in the liquid can be fixed on the surveyed area and precipitate.As shown in Figure 2; In another concrete embodiment; In order to practice thrift the specific bond molecule of band color mark material, on reagent areas 115, can also handle some colored dyestuffs (35), when detecting; Dyestuff in the dyestuff zone is taken on the testing result control area by liquid and is precipitated by flocculation reagent or coagulum, and so just Show Color representes that testing result is effective on the testing result control area.
Like Fig. 3-6; In other embodiments shown in Figure 8; On positive control area 55,455, can handle some flocculation reagent or coagulums; These flocculation reagent or coagulum can precipitate colloidal solid or dye granule in the liquid, no matter whether contain analyte in the sample, on positive control area, all can have color to occur.
The flocculation reagent of indication of the present invention or coagulum comprise the reagent that arrives commonly used in the wastewater treatment.Inorganic polymer flocculation reagent (IPF) and the organic polymer reagent that flocculates for example.Wherein inorganic polymer flocculation reagent mainly is to be many derivants that base stock prepares with the aluminium polychloride, like polymeric ferric aluminum, polymeric aluminium-silica and organic composite type flocculation reagent.
The most commonly used is organic polymer flocculation reagent, and organic polymer flocculation reagent has synthetic and natural two kinds.These flocculation reagent by the difference of functional group can the molecular cation type, nonionic and anionic.Water-solublely by whether can be divided into water-soluble and water-insoluble flocculation reagent.The for example polyacrylamide of nonionic (PAM), polyoxyethylene, anionic polypropylene acid (PAA), gather methyl-prop dilute acid hydrolysis polyacrylamide (HPAM), gather sulfo group styrene; Cationic is butyl bromide ethene pyrrole, gathers propyl-dimethyl amine (PDADMA).Organic polymer flocculation reagent has flco and the consequent huge surface adsorption effect that formation is bigger between particle.Contain in the water soluble organic polymer polymkeric substance of those high polymerization degrees or the molecule of multipolymer many can with glue particle and the fine particle suspended particle surface acting active group in some site, molecular weight hundreds thousand of to millions of.The nonionic functional group of flocculating polymer reagent be hydroxyl (OH), (CN) and acid amides (CONH2); Cationic functional group be primary amine (NH2), secondary amine (NH-R), tertiary amine (NH-RR) and quaternary amine (R-N-RR), anionic functional group such as carboxylic acid (COOH), sulfonic acid (SO3H) and sulfuric acid ester (OSO3H) or the like.
These flocculation reagent or coagulum are applied for publication 200510023481,200410024480,200310122818,00130879 and 01129968 in China, and in Chinese patent 99116150,98116440 and 86104833 detailed description are arranged all.These organic or inorganic flocculation reagent can be through buying to the blue ripple chemicals in Wuxi company limited.Of the present invention one preferred embodiment in, the flocculation reagent of use is water-soluble high analyte organic-flocculation reagent.
To the control area, can use the prior art known method to the flocculation agent treated.The reagent of these commercial uses can directly use, and it is better that general inorganic flocculation reagent can be regulated suitable pH value back result of use; To flocculating polymer reagent, in order to obtain better disperse state, ordinary priority should be selected for use and can in water, evenly disperse, dissolve, and has the macromolecular compound matter or the water-soluble high-molecular compound matter of adsorption activity group.In a concrete embodiment; Commercial wastewater treatment flocculation reagent commonly used can be diluted to certain concentration with pure water or soft water earlier; Regulating the Membrane jetter device with the trace of a microprocessor device control then is sprayed directly on on the nitrocellulose filter; Dry then, in addition, to nitrocellulose filter, also be fine with liquid-transfering gun manual process flocculation reagent.Upward the flocculation reagent of wastewater treatment also can be with being processed on the film after the certain concentration of some buffer solution dilutions again in these industry.Handled film is not had special requirement, all is feasible as long as can fix the film of some albumen, for example nitrocellulose filter, nylon membrane, CAM, filter paper or the like.The membrane material that any persons skilled in the art combine the present invention to expect.One preferred embodiment in, can also in flocculation reagent dilute solution, add some surface-active agents.
Reagent areas and specific bond molecule
The marked region that comprises the specific bond molecule that has mark substance on the reagent areas; Be fixed with another specific bond molecule on the surveyed area, the specific bond molecule that has mark substance can specific bond form a signal with another specific bond molecule on surveyed area.Produce signal and can be based on three sandwich method or the competition laws of showing one's high ideals, perhaps their other methods of deriving.The specific binding molecule of analyte be meant combine with analyte and can not with the binding molecule of other any molecule strong bonded in the sample.The specific binding molecule of analyte also can combine with existence that shows analyte in sample or the molecule that is associated with the existence of analyte.Strong bonded is meant to combine to reach and changes test findings or make the unconspicuous degree of test findings.In some concrete schemes; Specific binding molecule possibly be a kind of antibody or a kind of antibody fragment (for example; A kind of Fab district of antibody); A kind of antigen, a kind of acceptor of binding partner or the fragment of acceptor, it is right that perhaps biotin-streptomycete avidin combines the combination of a right composition or other type.Reagent area just can provide mark like this, and when sample flow was crossed reagent area, analyte had combined to produce the mark of detectable signal." label pad " is meant the position of the material that contains the analyte that possibly exist in the underlined sample on the matrix.Therefore a reagent area is exactly a label pad." mark " can be any suitable mark that produces detectable signal.For example, mark can be a sol particle, fluorescent grain, and chemiluminescent molecule, metal or alloy (for example, collaurum), perhaps capsule particularly comprises the liposome of visible dyes.Hydrophobic sol also is that useful, hydrophobic organic dyestuff or pigment are soluble or have only very limited sub-fraction solvable in water.Mark can also be a polymer particles, for example coloured granules of polystyrene (for example, spherical).Other useful granular mark comprises ferritin, phycoerythrin, phycobilin-albumen, metal or alloy deposition or solubility, fungi, marine alga, perhaps pigment of bacterium or derivant, for example chlorophyll of bacterium or other plant material.In some concrete scheme, mark is a coloured particle, for example dextran bead.In other concrete scheme, as the mark color identical of positive control with dye selection, the interaction when producing single tangible symbol on matrix or in the matrix to strengthen two kinds of signals.
In other concrete scheme, mark possibly be the specific binding molecules (for example, a kind of antibody) that analyte a kind of has been labeled.For example, in a concrete scheme, the target analyte is human chorionic gonadotrophin (hCG), is the anti-hCG antibody of aurosol mark in conjunction with the mark of hCG.When sample arrived reagent area (perhaps label pad), the hCG in the sample was by the anti-hCG antibodies of aurosol mark.Labelled antibody does not disturb the capture molecules of analyte land and the hCG of mark to combine.For example, mark can combine a part of analyte, and capture molecules can combine another part or the incorporation of markings of analyte.HCG-is anti--and hCG antibody-gold compound moves to the downstream of matrix.When compound arrives the analyte land with capture molecules combine form gold-anti--hCG anti--hCG-resists-hCG antibody.Capture molecules possibly be the another kind of specific binding molecules of hCG, or combines the specific binding molecules of the halfbody of hCG analyte.When gold-anti--hCG specific binding molecules-hCG-anti--when hCG specific binding molecules compound is attached to the analyte land, the analyte land by the golden marker coloring on the compound and in the analyte land gold mark naked eyes that become visible.In a concrete scheme, specific binding molecule is antibody or antibody fragment.Mark with catch binding molecule and can combine on the analyte different antigens decision position, in a concrete scheme, the specific binding molecules of mark has combined β-hCG, has combined α-hCG and catch binding molecule.
" antibody " is meant immunoglobulin (Ig), no matter is natural or some or all of synthetic.This term also comprises the derivant of the antibody that wherein keeps binding ability, also comprise any contain with the binding domain homologue of immunoglobulin (Ig) or the protein that combines the territory of homology to a great extent.These protein possibly be to be derived from natural materials, also possibly be some or all of synthetic.A kind of antibody possibly be monoclonal or polyclonal.A kind of antibody possibly be a member in any immunoglobulin class, comprises any mankind's immunoglobulin class: IgG, IgM, IgA, IgD, IgG and IgE." antibody fragment " is a part less than total length of the derivant or the antibody of antibody.Antibody fragment can remain to the remarkable site of the binding ability of a few full length antibody.The example of antibody fragment comprises Fab, Fab ', F (ab ')
2, scFv, Fv, dsFv dimer and Fd fragment, but not only comprise above these.
Antibody fragment can be generated by any way.For example, antibody fragment can generate through enzymolysis or complete antibody of chemical cracking, perhaps also can be through the genetic recombination from the coded portion antibody sequence.In other words, the antibody fragment generation of can partly or wholly recombinating.Antibody fragment can be a single chain antibody fragments arbitrarily.In other words, antibody fragment can comprise many peptide chains that interconnect, for example, and through disulfide linkages.Antibody fragment also can be a kind of arbitrarily polymolecular compound.One has the antibody fragment of function to comprise about at least 50 amino acid usually, and more antibody fragment comprises about at least 200 amino acid usually.
Strand Fvs (scFvs) is the antibody fragment of reorganization, and it is only by variable light chain (V
L) and variable heavy chain (V
H) each other with the polypeptied chain covalent bond.V
LAnd V
HIn a side have the amido end regions.Polypeptied chain length is variable with forming, and its length can make two mutual bridgings of variable domain and the arrangement of atom is not had a strong impact on.Polypeptied chain mainly is made up of glycocoll and serine residue extension usually, wherein has some glutamic acid and lysine residue to be dispersed in distribution to increase its solubleness." dimer " is meant the dipolymer of strand Fvs.The peptide chain that dimeric monomer comprises is usually than the weak point of most of strand Fvs, and they demonstrate the tendency that forms dipolymer.
" Fv " fragment is by a V
HWith a V
LThe territory is with the non-covalent composition that interconnects.Term " dsFv " here is meant and comprises a stable V
H-V
LThe Fv of right intermolecular disulfide bond." F (ab ')
2" fragment is a fragment of antibody, in essence with identical with the pepsin fragment that digestion immunoglobulin (Ig) (normally IgG) obtains when the pH4.0-4.5.This fragment also can be re-combined into." Fab ' " fragment is a kind of antibody fragment, in essence with through reducing F (ab ')
2The fragment that two interconnective cystine linkages of heavy chain on the fragment obtain is identical.Fab ' fragment also can be re-combined into.“Fab”
Fragment be a kind of in essence with the identical antibody fragment of fragment that obtains with papain digestion immunoglobulin (Ig) (usually IgG).The Fab fragment also can be re-combined into.Heavy chain fragment on the Fab fragment is the Fd fragment.
The said reagent areas of the present invention can comprise one or several reagent areas, and these reagent areas differ to establish a capital and exist on the reagent strip certainly.In one embodiment, reagent areas 115 is positioned at the upper reaches (like Fig. 1-8) of surveyed area 95.In a concrete embodiment; Reagent areas 115 can comprise a marked region 15 and dyestuff zone 35; On marked region, handle the specific bond molecule that band color mark material is arranged; On the dyestuff zone, handling has colored dyestuff, does not have the specific bond molecule to have (Fig. 2) on this dyestuff.In other embodiments; Also handle the chemical reagent that the conditioned reaction condition is arranged on the reagent areas; These chemical reagent condition that can offer the best for the reaction between specific bond molecule on the surveyed area and the band colored particle mark substance molecule; The pH value of conditioned reaction for example reduces the interfering material of other disturbance reponse in the sample.These chemical reagent can be handled in the sample region of acceptance, also can handle between dyestuff zone and surveyed area.
In another embodiment, reaction reagent can elder generation and sample mixing, and then is inserted into the reagent strip that contains surveyed area and control area and goes in the solution to react.
In another embodiment; On reagent areas, comprise marked region 15; On marked region, handle the excessive specific bond molecule that has mark substance is arranged; Like this when flow of liquid is crossed positive control area 55, a part of mark substance in the solution flocculated reagent or nonspecific the catching of coagulum and produce change color; On specific bond analyte zone, the specific bond molecule produces specific bond (Fig. 5) with the specific bond molecule that has mark substance.
The specific bond molecule and the mark substance of tape label material
The specific bond molecule of said here tape label material is meant that this molecule and another specific bond molecule that is fixed on the surveyed area are man-to-man combinations.It is that a molecule passes through physics or another molecule of chemical mode specific bond that so-called specificity combines, and mutually combining between these two molecules can combine difference mutually with other.Specific bond between this two molecules is except the antibody that comprises antigen and antigen; Another antibody that also comprises antibody and anti-this antibody; The albumen of biotin and antibiotin, between the polypeptide fragment, DNA and DNA; RNA and RNA, and pass through specific bond pairing molecule of recombinant technique acquisition or the like.This combination can be direct combination, can also be to close through the intermolecular access node of its special pairing.Specifically, the combination that is fixed between the molecule of specific bond molecule and tape label material on the surveyed area can detect antigen by double antibody sandwich method, and dual-antigen sandwich method detects antibody or derives other directly and indirect method by their institutes.These specific bond are that those skilled in the art combine the branch invention easily to arriving.
Here said mark substance is meant one type of material that can read through naked eyes Direct observation or machine.The mark substance of many types can produce through physics or chemical mode can identified signal.For example enzyme, fluorescent material, chemiluminescent substance, radiomaterial, other material comprise self band colored particle shape material, and this particulate material can be metallic colloid or latex colloid or the like, can also be water-soluble dye.
The type of sample and analyte
The sample of any kind can both make an experiment with device of the present invention, comprises body fluid (for example, urine and other body fluid, and clinical sample).Fluid sample possibly be derived from solid or semisolid sample, comprises excrement, biological tissue and foodstuff samples.These solids can be transformed into fluid sample through any suitable method with semisolid sample, for example in a kind of suitable liquid, mix, stamp broken; Macerate, hatch, dissolving or enzymolysis solid sample are (for example; Water, phosphate buffer or other damping fluid)." biological sample " comprises the sample that is derived from animal alive, plant and food; Also comprise urine, saliva, blood and blood constituent, cerebrospinal fluid, vaginal swab; The culture of seminal fluid, ight soil, sweat, secretion, tissue, organ, tumour, tissue and organ; Condition medium cell culture and there is no matter be the people's or animal.Foodstuff samples comprises finished COF and last product, meat, cheese, wine, milk and potable water.Plant sample comprises the sample of the condition medium that is derived from any plant, plant tissue, plant cell cultures and there." environmental sample " is those samples that are derived from environment (for example, the sample of lake water sample or other water body, sewage sample, pedotheque, underground water sample, seawater sample, the samples of refuse water).Sewage also can be included in the environmental sample with relevant refuse.
Can analyze any analyte with the present invention.The example of the analyte that the enough the present invention of ability detect comprises (but not only comprising) human chorionic gonadotrophin (hCG), lutropin (LH), ovarian stimulation plain (FSH); Hepatitis C virus (HCV); Hepatitis B (HBV), hepatitis B surface antigen, the medicine of AIDS virus and any abuse.Analyte can detect in any liquid or liquefied sample, urine for example, saliva, saliva, blood, blood plasma, perhaps serum.The example of other analyte also has the acid of flesh ammonia acid anhydride, cholerythrin, nitrite, protein (nonspecific), blood; Leucocyte, blood sugar, heavy metal and toxin, bacterium composition (for example, special protein and the sugar of the bacterium of particular type; Colon bacillus 0157: H7 for example, staphylococcus aureus, salmonella, C.perfringens; Campylobacter, listeria monocytogenes, enteritis vibrios, perhaps cured shape bacillus).Any other analyte of suitable lateral flow assay form can detect with this device.
Reagent strip and the device that has reagent strip
What said here reagent strip referred to can let liquid at the last reagent strip that flows, and on reagent strip, comprises surveyed area and control area at least.This reagent strip can be made by a lot of materials, and for example cellulose, filter paper, nitrocellulose filter or nylon membrane or the like are anyway as long as can let liquid just passable from the material that an end of reagent strip flows to the other end.
In a concrete embodiment, this reagent strip is the nitrocellulose filter reagent strip.Specifically, as shown in Figure 6, this reagent strip 6 is made up of four parts, comprises sample absorbing sheet 425, marker slip 4115 and cellulose nitrate sheet 475 and suction sheet 465.These several parts interconnect and can let liquid arrive suction sheet 465 from sample blank film 425 through marker slip 4115, nitrocellulose filter 475.Wherein, On marker slip, handle the specific bond molecule that has colored particle is arranged; On nitrocellulose filter 475, comprise surveyed area 495; Surveyed area 495 comprises the zone 445 and positive control area 455 of containing the specific bond analyte, on positive control area, is fixed with a certain amount of flocculation reagent or coagulum.The material that constitutes these sheets can some common absorbent materials, as spun glass is plain, polyester fiber is plain, filter paper or the like.In a preferred embodiment, the sample blank film is that the plain sheet of spun glass, marker slip are that polyester diaphragm, suction sheet are filter paper.In another embodiment, be to also have a testing result control area 485, on surveyed area, be fixed with a certain amount of flocculation reagent or coagulum (Fig. 4) in the downstream of surveyed area with embodiment difference shown in Figure 6.In any one embodiment of describing in the above, the part of forming reagent strip can be fixed on the hydrophobic backing plate.In another embodiment; This reagent strip can be placed in the device that contains upper plate and lower plate and go; Having the sample region of acceptance on a liquid through-hole and the reagent strip to communicate on the upper plate of this device, it is corresponding that the result of upper plate reads window and surveyed area and testing result control area.The material of these reagent strips and device are that those skilled in the art combine the present invention to expect easily, and they are not emphasis of the present invention, and the material of formation reagent strip various piece and the device that holds reagent strip are all in United States Patent (USP) 5,602,040; 5,989,921; 6,352,862; 6,319,676,5,824,268; 6,184,040; 6,472,160; 6,267,722; 6,649,418; 6,537,828; Detailed announcement is all arranged in 5,739,041.
For the beneficial effect of the invention better is described, give description of test at present
The detector bar that experiment 1 usefulness has the reagent that flocculates detects human chorionic gland promotion hormone (HCG)
This example has been described a concrete scheme using reagent strip of the present invention to detect the HCG in the urine
The preparation of reagent.Flocculation agent treated: with distilled water dilution BWD-Q1 decolouring flocculation reagent (the blue ripple chemicals in Wuxi company limited) and add the surfactivity preparation of S-10.The ratio of dilution is followed successively by (flocculation reagent: (volume ratio): 1:50 water), 1:100,1:200,1:400,1:800,1:1600.
Dyestuff is handled: use pH is 7 PBS buffering liquid dilution eosin W or W S dyestuff (reagent company limited is thought carefully in Shanghai).The final concentration of dilution is: 0.2%, 0.1%, 0.055%, 0.025%, 0.0125%, 0.006255%.
Reagent strip of the present invention is prepared.Reagent strip is according to commonsense method manufacturing in this area.With reference to Fig. 6; On marker slip 4115, handle mouse anti β hCG IgG and the eosin W or W S dyestuff that contains the aurosol mark; On the analyte calmodulin binding domain CaM 445 of nitrocellulose filter 475, handle goat-anti α HCG IgG (4.0mg/ml), treatments B WD-Q1 decolouring flocculation reagent on the positive control area 455 of nitrocellulose filter 475.Can be sprayed at these agent treated on the film arbitrarily with the micro-flusher of a microprocessor control on the nitrocellulose filter.Have sheet material (4115, the 475) drying of having handled well to these then.After dry getting well, sample blank film 425, marker slip 4115, nitrocellulose filter 475 and suction sheet 465 are connected in turn according to shown in Figure 6.The concentration of the BWD-Q1 decolouring flocculation reagent on dye strength of handling on the marker slip and the positive control area is come the reagent treatment bar according to the combination shown in the table 1.
The preparation of contrast agents bar.With the difference of reagent strip of the present invention is exactly to handle on the positive control area of contrast agents bar anti-human IgG (1.3mg/ml) is arranged, and on reagent areas, does not have dyestuff.Other all the same.
Detect negative and positive urine liquid.Reagent strip according to above processing detects feminine gender (10) and positive urine liquid (10) respectively, does control experiment simultaneously.
Experimental result 1:
Table 1: the control lines experimental result of the different B WD-Q1 decolouring flocculation reagent concentration on the control area
Experimental result 2:
Table 2: the flocculation reagent of handling on the control area and the specific antibody of processing are to the influence of testing result
Conclusion
From the result shown in the table 1, the flocculation reagent of on positive control area, handling can reach the result of G8-G9, when detecting, very bright-coloured red lines on positive control area, occur.The background of this reagent strip was fine when dye reagent concentration was 0.0125%-0.00625%, and is bright-colored.
Simultaneously, can find out from table 2 that we detect the positive urine fluid samples with these reagent strips, detect the result who all shows positive sign "+" on the control area, negatives all demonstrates negative sign "-" result, and is consistent with the testing result of the reagent strip that contrasts.Then explanation, BWD-Q1 on this positive control area decolouring flocculation reagent does not influence the specific binding reaction on the analyte calmodulin binding domain CaM.
Here G8, G9 are the internal control standards of weighing shade, generally are to deepen gradually from colourless to the G12 color from GO, and general is just can reach the requirement of commercial product more than the G7 in color on the control area.The depth of background also is a control of product standard; The reason that produces background be these colored particles (dyestuff and mark substance) on nitrocellulose filter because capillary layer is inhaled forward flows; These particles are on film owing to will receive the resistance of the micropore on the nitrocellulose filter and show color belt; Along with flowing of liquid, the disappearance that these colors can be gradually.General coml requires bright-colored for what occur on the control area, and the speed that background shoals in certain hour is fast more, and then the quality of product is just good more.
The invention that this paper describes for example just can be used under the situation that each part all possesses, and being limited in here of it just do not specified.It is not unique constant being used for the term and the expression way of tracing device among this paper; And the expression way of the structure that we have no intention to use these terms and expression way to get rid of to describe this device or any same meaning of characteristic, we admit the various expression way in the scope of claim of the present invention.Therefore; Although we think the present invention in this article with various concrete schemes and arbitrarily feature description clearly display; But the expression way of the design that discloses among change this paper also will be sought help from those experienced professional technique personages, and these changes are consistent with the claim that the present invention attaches.
Useful digitized information that mention and that quote as proof combines among the content of article, patent, patent application and all other documents and this paper; Must come reference as a complete content, delivering wherein, any one part all will specialize this point.The applicant has these any and whole articles, patent, patent application or the information of other document and the right that material is integrated with this application book.
Claims (10)
1. whether contain the pick-up unit of analyte in the test sample, comprising: support the reagent strip that liquid sample flows, on reagent strip, comprise surveyed area, control area and sample receiving area; Surveyed area comprises the analyte calmodulin binding domain CaM; This calmodulin binding domain CaM comprises a species specific binding molecule, and this device also comprises and is positioned at downstream, sample receiving area, the reagent areas at the surveyed area upper reaches; Has colloidal solid in this reagent areas; Dyestuff or water-soluble mark substance is characterized in that, comprise non-albumen flocculation reagent on the described control area.
2. pick-up unit as claimed in claim 1 is characterized in that, described reagent strip is the nitrocellulose filter reagent strip, wherein, is followed successively by sample receiving area, reagent areas and surveyed area along being from upstream to downstream on the liquid flow direction.
3. pick-up unit as claimed in claim 2 is characterized in that described reagent areas also comprises marked region.
4. pick-up unit as claimed in claim 3 is characterized in that, described control area is the testing result control area, and this testing result control area is positioned at the downstream of surveyed area.
5. pick-up unit as claimed in claim 2 is characterized in that, described control area comprises testing result control area and positive control area, all includes described non-albumen flocculation reagent on testing result control area and the positive control area.
6. pick-up unit as claimed in claim 5 is characterized in that the testing result control area is positioned at the downstream of surveyed area, and the analyte calmodulin binding domain CaM on positive control area and the surveyed area intersects to form the symbol that can discern.
7. pick-up unit as claimed in claim 6 is characterized in that, described positive control area and described analyte calmodulin binding domain CaM intersect to form a crux.
8. like claim 3 or 5 described pick-up units, it is characterized in that described reagent areas also comprises marked region.
9. like claim 3 or 5 described pick-up units, it is characterized in that described reagent areas also comprises a dyestuff zone, on the dyestuff zone, comprise a certain amount of dyestuff that has color, this dyestuff zone is positioned at the upper reaches of surveyed area.
10. like claim 1, one of 5 or 6 described pick-up units, it is characterized in that described non-albumen flocculation reagent is water soluble organic polymer flocculation reagent.
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| US4916056A (en) * | 1986-02-18 | 1990-04-10 | Abbott Laboratories | Solid-phase analytical device and method for using same |
| CN1342902A (en) * | 2000-09-11 | 2002-04-03 | 同济医科大学 | Test paper for quickly diagnosing Schistosomiasis japonica |
| US6855561B2 (en) * | 2001-09-10 | 2005-02-15 | Quidel Corporation | Method for adding an apparent non-signal line to a lateral flow assay |
| CN1682114A (en) * | 2002-06-27 | 2005-10-12 | 因韦尔尼斯医药瑞士股份有限公司 | Assay device for liquid sample |
| CN2938100Y (en) * | 2006-04-12 | 2007-08-22 | 艾博生物医药(杭州)有限公司 | Reagent strip containing non-protein trapping matter at control area |
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| US4916056A (en) * | 1986-02-18 | 1990-04-10 | Abbott Laboratories | Solid-phase analytical device and method for using same |
| CN1342902A (en) * | 2000-09-11 | 2002-04-03 | 同济医科大学 | Test paper for quickly diagnosing Schistosomiasis japonica |
| US6855561B2 (en) * | 2001-09-10 | 2005-02-15 | Quidel Corporation | Method for adding an apparent non-signal line to a lateral flow assay |
| CN1682114A (en) * | 2002-06-27 | 2005-10-12 | 因韦尔尼斯医药瑞士股份有限公司 | Assay device for liquid sample |
| CN2938100Y (en) * | 2006-04-12 | 2007-08-22 | 艾博生物医药(杭州)有限公司 | Reagent strip containing non-protein trapping matter at control area |
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