CN101017169A - Analysis equipment of biological sample - Google Patents
Analysis equipment of biological sample Download PDFInfo
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- CN101017169A CN101017169A CN 200610052628 CN200610052628A CN101017169A CN 101017169 A CN101017169 A CN 101017169A CN 200610052628 CN200610052628 CN 200610052628 CN 200610052628 A CN200610052628 A CN 200610052628A CN 101017169 A CN101017169 A CN 101017169A
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Abstract
This invention relates to one device to test liquid sample, which comprises agent bar and second agent area, wherein, the agent bar comprises sample receive area and label area first agent area and test area; the test area comprises certain abnormal combination molecule composed of the analysis mass irrelative abnormal molecule and other molecule; the second agent area is on upstream of the test bar in discrete flow status; the second agent area comprises abnormal combination molecule on the analysis subject and the abnormal combination module irrelative abnormal property.
Description
Field that the present invention belongs to
The present invention relates to sample testing apparatus, more specifically, be meant a kind of detection system that detects low concentration small-molecule substance in the sample that is applicable to.
Description of related art
The device that utilization contains reagent strip detects and whether contains analyte in the sample and all having in a lot of prior aries and disclose or describe.For example shown in Fig. 1 C, these reagent strips 10 generally comprise reagent areas and surveyed area 13, and wherein reagent areas can comprise sample region of acceptance 11 and marked region 12.On surveyed area 13, comprise zone 132 that shows testing result and the testing result control zone 133 that is positioned at the surveyed area downstream.Usually, the sample region of acceptance of these reagent strips comprises sample blank film 111, marked region comprises marker slip 121, reagent areas is formed in these two zones jointly, on reagent areas, comprise and finish the necessary reagent of reaction, surveyed area comprises detection lug 131, the testing result control control zone 133 that on detection lug 131, comprises the zone 132 that shows test results and be positioned at these 132 downstreams, zone, general testing result zone is a linear, and Show Color changes and represents whether analyte exists in the sample on the testing result zone; Suction zone comprises suction sheet 141, and these parts 111,121,131,141 join end to end and allow liquid can the direction from an end of reagent strip along the arrow indication flow to the other end to finish test.General commonly used reagent strip is the nitrocellulose filter reagent strip, and promptly surveyed area comprises nitrocellulose filter, and fixedly the specific bond molecule shows the result of detection on nitrocellulose filter; Can also be cellulose acetate membrane or nylon membrane or the like.The reagent strip that for example the following patent is described or contain the device of reagent strip: US 4857453; US 5073484; US 5119831; US 5185127; US 5275785; US 5416000; US 5504013; US 5602040; US 5622871; US 5654162; US 5656503; US 5686315; US 5766961; US 5770460; US 5916815; US 5976895; US 6248598; US 6140136; US 6187269; US 6187598; US 6228660; US 6235241; US 6306642; US 6352862; US 6372515; US 6379620; With US 6403383.
Utilize these pick-up units that contain reagent strip to detect the method that whether contains analyte in the sample and generally include competition law and non-competing method, non-competing method comprises that the method that double antibody or double antigens sandwich or its are derived detects antigen or antibody, utilizes competition law generally to be used to detect the haptens small-molecule substance.But, when containing the lower analyte of concentration ratio in the sample, utilize the pick-up unit of these prior aries to detect the complicated operation that just seems, and testing result is not very desirable, thereby may produces some inaccurate results.In addition, when utilizing the pick-up unit of prior art to carry out the detection of analyte in the sample, usually in addition need to detect buffering liquid and just can finish detection, complicated operation and may produce other adverse influence to the operator like this is such as the safety issue of buffering liquid etc.The present invention provides a kind of new pick-up unit in order to overcome these deficiencies of prior art, not only can improve the sensitivity of detection, simultaneously, when the sample that detects is difficult to handle, can also alleviate the loaded down with trivial details degree of operation greatly, improve the efficient and the accuracy that detect.
Summary of the invention
The present invention relates to a kind of sample testing apparatus, comprise reagent strip and second reagent areas of separating with reagent strip in the device, the reagent strip and second reagent areas are in the liquid communication state; Reagent strip comprises first reagent areas and surveyed area.When detecting, tracer liquid is through second reagent areas and then flow to first reagent areas on the reagent strip, thereby finishes detection reaction.Utilize this device not only can improve the sensitivity of detection, in addition, it is just passable to utilize device of the present invention sometimes only to need to collect sample, need not also need to add extra buffer solution and finish reaction as traditional gathering-device, for example special the interpolation extracted buffering liquid for the small-molecule substance in the separating sample.When having the small-molecule substance of low concentration in the detection sample, pick-up unit of the present invention is effective especially especially.
On the one hand, the invention provides a kind of pick-up unit that whether has analyte in the sample that detects, device comprises reagent strip, reagent strip comprises first reagent areas and surveyed area, on surveyed area, comprise fixed specific bond molecule, first reagent areas is positioned at the upstream of surveyed area, this device also comprises second reagent areas of separating with reagent strip, this second reagent areas comprises can be with the specific bond analyte molecule of liquid flow, and second reagent areas is positioned at the upstream of first reagent areas and is in the liquid communication state with first reagent areas.When detecting, if the analyte in the sample exists, they just with second reagent areas on specific bond analyte molecule react, flow on the reagent strip with sample then, finish reaction at surveyed area.Utilize sensitivity that this device can be higher to detect the analyte of low concentration in the sample, be particularly suitable for detecting the small-molecule substance of low concentration in the sample.
Detect analyte in the sample based on non-competing method, in examples of implementation, comprise on second reagent areas in the device a kind of can with the molecule (Y1) of the specific bond analyte of liquid flow and with the incoherent specific bond molecule of analyte to a kind of molecule (M1) in (M1/M2), first reagent areas on the reagent strip also comprises marked region and sample region of acceptance, comprise on the marked region that the another kind that links to each other with mark substance (L) can be with the molecule (Y2) of the specific bond analyte of liquid flow, on surveyed area, comprise with the incoherent specific bond molecule of analyte to the another kind of molecule (M2) in (M1/M2), this molecule is fixed on the surveyed area.When detecting, sample at first with device in second reagent areas on reagent react, if there is analyte (A) in the sample, the molecule of specific bond analyte just and analyte in conjunction with formation compound substance (M1-Y1-A); Sample flows on the marked region then, and the molecule (Y2) of the another kind of specific bond analyte on the marked region also and analyte (A) combination; When the compound substance (M1-Y1-A-Y2-L) of their formation is on the process surveyed area at last, on the surveyed area and another kind of molecule (M2) the incoherent specific bond molecule of analyte centering show the result of test with regard to these compound substances of specific bond on surveyed area.The A here, Y1, Y2, L, M1 and M2 are on adding for convenience of understanding, and can not constitute the restriction to these schemes.In specific embodiment, dual-antigen sandwich method, when analyte is antibody, a species specific antigen that comprises this antibody correspondence on second reagent areas, this antigen with link to each other or combination with a kind of molecule of analyte (antibody) incoherent specific bond molecule centering, on marked region, comprise colored mark substance, colored mark substance links to each other or combination with another corresponding specific antigen of analyte (antibody), comprises fixed on surveyed area and the another kind of molecule of the incoherent specific bond molecule of analyte (antibody) centering.When detecting, sample to be detected at first with second reagent areas on reagent reacting, flow to marked region and surveyed area on the reagent strip then, if contain analyte in the sample, color appears on surveyed area, then represent positive findings; If there is not analyte in the sample, on surveyed area change color can not take place, then be expressed as negative findings.In another concrete scheme, double antibody sandwich method, analyte can be antigen, on second reagent areas, comprise this antigen of specific bond a kind of antibody and with a kind of molecule of the incoherent specific bond molecule of this antigen centering; On marked region, comprise and linking to each other with mark substance or the antibody of the another kind of specific bond analyte of combination; Another kind of molecule in fixing and the incoherent specific bond molecule of analyte antigen on surveyed area.Utilize these devices, when in sample, containing the very low analyzed material of concentration, at first this sample can fully contact with reagent on second reagent areas and react the suitable time, and then allows sample flow to finish reaction on the reagent strip, can improve the sensitivity of detection so greatly.Particularly when the detection sample is saliva sample, utilize this pick-up unit more favourable, because it is more much lower than the concentration in other body fluid to contain analyte of interest matter in the saliva, second reagent areas reaction in the increase saliva in analyte of interest matter and the device, the influence of other has been handled on the reagent strip reagent can be reduced like this, thereby the susceptibility of detection can be improved greatly sample.
Detect analyte in the sample based on competition law, in another examples of implementation, comprise on second reagent areas specific bond analyte molecule (Y1) and with the incoherent specific bond molecule of analyte to a kind of molecule (M1) in (M1/M2), reagent strip comprises first reagent areas, first reagent areas comprises sample region of acceptance and marked region, comprise that on marked region another that link to each other with mark substance (L) can be with the similar substance (A*) of the analyte of liquid flow, comprise on surveyed area and the another kind of molecule (M2) of one of the incoherent specific bond molecule of analyte centering that this molecule (M2) is fixed on the surveyed area.When detecting, when the detection sample that contains analyte passes through, flows through or passes through second reagent areas or after stopping certain hour on second reagent areas, analyte in the sample, if have, the molecule of specific bond analyte is elder generation and analyte specific bond formation compound just; When these compounds, analyte (if also having) or remaining specific bond analyte molecule (if residue is arranged) just flow to first reagent areas on the reagent strip with sample together, with regard to and on the first reagent areas marked region, react.Come Show Color to change in the expression sample whether have analyte by surveyed area then.
In a concrete embodiment, when utilizing competition law to detect in the sample small-molecule substance, on second reagent areas, handle the antibody of the analyzed small-molecule substance of certain density specific bond, on antibody, also will connect a kind of molecule in advance with the incoherent specific bond molecule of analyte centering; The necessary reagent of processing reaction on the sample region of acceptance on the reagent strip, for example the buffer solution of conditioned reaction pH value or other chemical reagent are handled the certain density similar substance that contains the analyte of labeled molecule on marked region.In the concentration of the antibody of second reagent areas and marked region and similar substance along with the standard that detects or require different and different, can regulate arbitrarily, the adjusting of these concentration is that one of ordinary skill in the art does not need performing creative labour to finish easily and to realize.For example, when analyzed micromolecule concentration is greater than certain concentration (C) in sample is set, the color showing positive findings on surveyed area, just do not occur,, the color showing negative findings just occurs when analyte the time less than the concentration (C) that is provided with.
In another examples of implementation, pick-up unit of the present invention comprises a body and collection well, and reagent strip is arranged in body, and collecting chamber comprises second reagent areas, and the reagent strip in this second reagent areas and the body is in the liquid communication state.Can also comprise on the body that testing result reads window and sample ingress area, testing result reads the surveyed area correspondence on the reagent strip in window and the body, and first reagent areas on sample ingress area and the reagent strip is in the liquid communication state.When detecting, sample is collected in the collecting chamber, allows the second reagent areas contact reaction in sample and the collecting chamber, allows sample flow to reagent strip from collecting chamber by the sample ingress area then and finishes whole detection reaction.In preferred embodiment, fix a kind of molecule on the surveyed area on the reagent strip with the incoherent specific bond molecule of analyte centering, first reagent areas on the reagent strip comprises can be with the mark substance of liquid flow, the similar substance of this mark substance and analyte links to each other or combination, second reagent areas that is arranged in collecting chamber comprise the other analyte molecule of a kind of specific bond and with the incoherent specific bond molecule of analyte centering another kind of molecule.In the tracer liquid sample, second reagent areas elder generation in the collection well and suitable time of liquid sample reaction, the liquid in making collecting chamber flows on the reagent strip by the sample ingress area and reacts then.In another preferred embodiment, comprise the antibody of specific bond analyte on second reagent areas in the pick-up unit collecting chamber and link to each other with this antibody or combination and a kind of molecule incoherent specific bond molecule of analyte centering, on marked region, comprise colored mark substance, the analog of colored mark substance and analyte in conjunction with or link to each other the another kind of molecule of the fixing and incoherent specific bond molecule of analyte centering on surveyed area.When detecting, sample collection, dropping, add or put in the collecting chamber, sample contacts with second reagent areas fully in collecting chamber and reacts, allow sample flow into then and finish whole detection reaction on the reagent strip, observe the change color of surveyed area by the window on the body and come whether to exist in the judgement sample analyte.
On the other hand, the present invention also provides the method that detects analyte in the sample.The method that whether has analyte in a kind of tracer liquid sample, comprise a kind of pick-up unit is provided, this pick-up unit comprises the reagent strip and second reagent areas, described reagent strip comprises first reagent areas and surveyed area, wherein first reagent areas comprises sample region of acceptance and marked region, and surveyed area comprises a kind of fixed specific bond molecule; Described second reagent areas comprise a kind of can be with the molecule of the specific bond analyte of liquid flow, and described second reagent areas is positioned at the upstream of sample region of acceptance on the reagent strip, and and described reagent strip be in the liquid communication state; Add the annex solution sample body on second reagent areas, this liquid sample flows on the described reagent strip by second reagent areas; By observing the result that the surveyed area change color shows detection.
On the other hand, the present invention also provides the kit of collecting and detecting sample, comprise sample testing apparatus and gathering-device, sample testing apparatus comprises reagent strip, on this reagent strip, comprise first reagent areas and surveyed area, wherein, described surveyed area comprises a kind of fixed specific bond molecule, first reagent areas is positioned at the upstream of surveyed area, described pick-up unit also comprises second reagent areas of separating with described reagent strip, this second reagent areas comprise a kind of can be with the molecule of the specific bond analyte of liquid flow.Gathering-device comprises collects handle and spongy top.The instructions that can also comprise operation in the kit.
Description of drawings
The pick-up unit that Fig. 1 describes prior art comprises the synoptic diagram of reagent strip.
Fig. 2 is the synoptic diagram of pick-up unit of the present invention
Fig. 3 detects preceding and detection back result schematic diagram for the pick-up unit of a concrete scheme of the present invention.
Fig. 4 detects preceding and detection back result schematic diagram for the pick-up unit of another concrete scheme of the present invention.
Fig. 5 detects preceding and detection back result schematic diagram for the pick-up unit of another concrete scheme of the present invention.
Fig. 6 is the pick-up unit synoptic diagram of specific embodiment of the present invention.
Fig. 7 is that the position of first gathering-device and reagent strip 2011 concerns synoptic diagram.
Fig. 8 is pick-up unit assembling diagrammatic cross-section.
Fig. 9 is the diagrammatic cross-section of pick-up unit before detection
Figure 10 and 11 is the diagrammatic cross-section of pick-up unit after detection.
Description of reference numerals
10,101,10-1,10-2 reagent strip; 11 sample regions of acceptance; 12 marked regions; 13 surveyed areas; 14 suction zone; 111 sample blank films; 121 marker slips; 131 detection lugs; 132 show the testing result zone; 133 testing result control areas; 141 suction sheets; 151 support chips; 201 second reagent areas; 30 pick-up units; 301 gathering-devices; 3011 collect handle; 3012 collect head; 308 collecting chambers lid; 302 first collecting chambers; 303 second collecting chambers; 601 collecting chambers; 305,305-1,305-3,305-2 O-ring seal; 306 sample ingress areas; 302-1,302-2,302-3,303-1,303-2,303-3 liquid through-hole; 306-1, the 306-3 sample imports through hole; 306-4 ingress area baseplane; 2011 reagent strips; 2012 paste-sides; 2013 reagent ends; The 306-2 snap ring; 3025 outer snap rings; 304 bodies; 3041 read window; The 306-3 sample imports through hole; Confirm chamber 307; 3071 bases; Derive through hole 3073; Stopper 3072; 3043 upper plates; 3045 lower plates.
Describe in detail
Come some embodiments of the present invention is described in detail below in conjunction with concrete accompanying drawing.These specific embodiment only are limited the enumerating under spirit of the present invention, do not get rid of one of ordinary skill in the art prior art and the present invention in conjunction with and other specific embodiments of producing.
Definition
Unless otherwise defined, all technology have identical implication with scientific terminology with the employed term of the those skilled in the art of this technical field that the present invention belongs to as used herein.
" detection " expression chemical examination or test a kind of material or whether material exists, such as, but be not limited to this, the metabolin of chemical substance, organic compound, mineral compound, metabolism product, medicine or drug metabolite, organic organization or organic organization, nucleic acid, protein or polymkeric substance.In addition, detect the quantity of expression test substances or material.Furtherly, immune detection, chemical detection, enzyme detection etc. are also represented in chemical examination.
" sample " refer to need chemical examination whether to exist and (or) analyze any material of analyte concentration, maybe need to determine whether one or more samples exist and (or) material of the analyte of quantity, maybe need to carry out the material of qualitative evaluation.Sample can be liquid sample, for example liquid sample.Liquid sample comprises body fluid, such as blood, serum, blood plasma, saliva, urine, tears, seminal fluid and marrow; Sample can be a water sample, such as seawater, river, river etc., perhaps from domestic water, municipal water use or service water resource, runoff water or sewage; Sample can be a food sample, such as milk and wine.Mucus, semisolid or solid sample can be used for making liquid, eluate, suspending liquid or leachate equal samples.For example, throat or genitals sample may be dipped in and make sample in the liquid.Sample can comprise potpourri or any relevant potpourri of liquid, solid and gas, such as the cell suspending liquid in dilution or the solution.Sample comprises biological substance, such as cell, microorganism, organelle and biochemical complexes.Liquid sample can be from producing such as solid, semisolid or high-viscosity materials such as soil, ight soil, tissue, organ, biological fluid or other occurring in nature non-liquid samples.For example, these solids or semi-solid samples can be mixed with the suitable solution of dilution one class.Sample can be macerated, freezing and thaw, perhaps other extracting method form liquid samples.Remaining particulate material can use traditional method such as filtration or precipitation to remove.
" upstream ", " downstream " refer to be divided along liquid flow direction, the upstream is to be positioned on the liquid flow direction, the downstream is positioned under the liquid flow direction, and upstream and downstream is a relative notion, and liquid can flow to downstream position from upstream position.
Reagent strip
General reagent strip, shown in Fig. 1 C, reagent strip 10 comprises that first reagent areas and surveyed area 13, the first reagent areas that are positioned at the first reagent areas downstream comprise sample region of acceptance 11 and marked region 12, surveyed area comprises a kind of specific bond molecule.More the reagent strip of You Huaing also comprises the suction zone 14 that is positioned at the surveyed area downstream, and sample can flow on reagent strip along the direction of arrow indication.On the sample region of acceptance, comprise and finish reaction necessary material, for example some buffer reagents, pre-service sample reagent or the like; Comprise the fluorescence labeling material at marked region, the colloid gold label particle, or with painted gel marking particle, it can also be water-soluble mark substance, these mark substances can connect antibody, antigen, haptens or analyte similar substance or the like, on surveyed area, comprise fixed specific bond molecule, change color can on surveyed area, occur by the specific bond molecule and represent whether there is analyte in the sample.Downstream at surveyed area can also comprise testing result control area 133.Reagent strip can be made up of following parts, and sample receiving pad 111 and label pad 121 are formed first reagent areas, and detecting pad 131, adsorptive pads 141, these materials are assembled into forms whole reagent strip 101 (as Figure 1B) on the support chip 151; Wherein comprise surveyed area 132 on detecting pad 131, preferential scheme is also to comprise testing result control area 133 in the downstream of surveyed area.Liquid sample can arrive adsorptive pads through label pad, detecting pad from the sample receiving pad in turn at last along the direction of arrow indication.Each parts of forming reagent strip can be to be made of absorbent material, and usually, detecting pad can constitute for one of nitrocellulose filter, cellulose acetate membrane or nylon membrane.These constitute the parts and the material of reagent strips and handle some common chemical reagent and disposal route all is that prior art is disclosed on these parts.
Be used for disclosed reagent strip and can be any known reagent strip of prior art, such reagent strip includes, but are not limited to: known immunoassays in the prior art, and the test of chemical assay and enzyme, such as, but be not limited to, the monospecific antibody immunoassays, multiple antibody mediated immunity is measured, and complex immunity is measured, the immunoassays of contrast, immunoassays of non-contrast or the like, comprise and utilize horseradish peroxidase, alkaline phosphatase, luciferase, the antibody pairing, antibody fragment, fluorescent-labeled antibody, modified antibody, labelled antibody, the antibody of colloid gold particle, or with antibody of painted gel particle mark or the like, they all are known in the prior art.The example that can be included into some reagent strips of this device can find in the United States Patent (USP) below: US 4857453; US 5073484; US 5119831; US 5185127; US 5275785; US 5416000; US 5424193; US 5504013; US 5602040; US 5622871; US 5654162; US 5656503; US 5686315; US 5766961; US 5770460; US 5916815; US 5976895; US 6248598; US 6140136; US 6187269; US 6187598; US 6228660; US 6235241; US 6306642; US 6352862; US 6372515; US 6379620; US 6403383; US 6656744; US 6979576; US 6372513; US 6372513.Further, the example that can be included into some reagent strips of this device can find in the U.S. Patent application below: sequence number is 09/579672; 09/579673; 09/653032; 60/233739; 09/915494,10/211199 and 09/860408.
Second reagent areas
Second reagent areas comprises can be with the molecule of the specific bond analyte of liquid flow, and this reagent areas is positioned at the upstream of reagent strip and separates with reagent strip.A scheme as shown in Figure 2, second reagent areas 201 is positioned at the upstream of the sample region of acceptance 111 of reagent strip 101, and liquid sample flows through the sample region of acceptance 111 that second reagent areas arrives on the reagent strip then earlier finishes whole detection.In specific embodiment, can also comprise on second reagent areas with the incoherent specific bond molecule of analyte to it a kind of molecule M1, the molecule Y1 of this molecule and specific bond analyte links to each other or combination, and this reagent areas can be positioned at the upstream of first reagent areas on the reagent strip.More specifically, second reagent areas is positioned at the upstream of the sample region of acceptance 11 of first reagent areas exactly, and sample region of acceptance 11 is positioned at the upstream of marked region 12, and marked region 12 is positioned at the upstream of surveyed area 13; On surveyed area, be fixed with the another kind of molecule Y2 of specific bond analyte, on marked region 12, comprise mark substance L and with the incoherent specific bond molecule of analyte to another kind of molecule M2 (Fig. 3 A).When reacting, if contain analyte A in the sample, then the specific bond molecule Y1 on the analyte A and second reagent areas carries out combination and forms compound substance A-Y1-M1; When this compound substance arrives on the marked region 12 by sample region of acceptance 11, just form new compound substance A-Y1-M1-M2-L; When new compound substance flows through on the surveyed area 13, be fixed on the new compound substance of specific bond molecule Y2 specific bond on the surveyed area, thereby color showing positive findings (Fig. 3 B) on surveyed area, occurs.
In another specific embodiment, can also comprise on second reagent areas with the incoherent specific bond molecule of analyte to it a kind of molecule M1, the molecule Y1 of this molecule and specific bond analyte links to each other or combination, and this reagent areas can be positioned at the upstream of first reagent areas on the reagent strip.More specifically, second reagent areas can be positioned at the upstream of the sample region of acceptance 11 of first reagent areas exactly, and sample region of acceptance 11 is positioned at the upstream of marked region 12, and marked region 12 is positioned at the upstream of surveyed area 13; On surveyed area, be fixed with the incoherent specific bond molecule of analyte to another kind of molecule M2, on marked region 12, comprise mark substance L and with the another kind of molecule Y1 (Fig. 4 A) of specific bond analyte.When reacting, if contain analyte A in the sample, then the specific bond molecule Y1 on the analyte A and second reagent areas carries out specific bond and forms compound substance A-Y1-M1; When this compound substance arrives on the marked region 12 by sample region of acceptance 11, just form new compound substance M1-Y1-A-Y2-L; When new compound substance flows through on the surveyed area 13, be fixed on the surveyed area with the incoherent specific bond molecule of analyte to the new compound substance of another kind of molecule M2 specific bond, form M2-M1-Y1-A-Y2-L, thereby color showing positive findings (Fig. 4 B) on surveyed area, occurs.
The incoherent specific bond molecule of analyte comprises (M1/M2), but be not limited only to this, biotin/affinity element (biotin/avidin), biotin/Streptavidin (biotin/streptavidin), antibody/antigen (antibody/antigen) (antibody and the analyte itself that do not comprise anti-analyte), the antibody of rhodamine/rhodamine (rhodamine/anti-rhodamine), antibody of mouse IgG/ mouse IgG (Mouse IgG/anti-mouse IgG) or the like.Analyte A can be antibody or antigen, and Y1 and Y2 can be specific antigen or the antibody molecule or the antibody fragments of different loci correspondence on the analyte.Mark substance L includes but are not limited to this, colloid gold particle, and latex particle, water-soluble mark substance or the like, mark substance is with some special antigens, antibody or other specific bond molecules link to each other or the technology of combination is existing technique known.Certainly, can also comprise other chemical reagent of some conditioned reaction system conditions in the sample region of acceptance, for example phosphoric acid buffer reagent, borate buffer reagent, carbonic acid buffer reagent or other albumen, big molecule or the like come optimizing reaction system.All to be persons skilled in the art expect and implement in conjunction with the present invention and prior art these optimized Measures easily.
The device that mask body is executed in the example in the utilization can detect low concentration or small-molecule substance in the sample, because the concentration of some important analytes in the sample very molecular weight of the end or analyte is very little, utilize traditional pick-up unit often can not detect or arrive requirement on the clinical meaning because the analyte concentration in the sample is very low.When particularly ought detect the analyte in some special analytes or some special samples of needs detection, traditional pick-up unit more can not satisfy accurately simply requirement fast again, for example detect certain small-molecule substance in the saliva, when utilizing traditional pick-up unit (Fig. 1) to put into sample in the pick-up unit through being everlasting, want simultaneously in addition in pick-up unit, for example on the sample region of acceptance 111, adding some reaction reagents or solution extracts, little amalyzing substances in the dissolving saliva just can be finished the requirement and the purpose of detection, before perhaps utilizing pick-up unit to detect, earlier the sample that will detect is carried out some processing, for example carry out dissolution process with chemical reagent solution, the filtration that precipitates with physical method or get rid of the interfering material that other may influence testing result.Especially when detecting sample and be saliva, want elder generation handle usually, allow be included in analyte in the saliva and discharge again and detect saliva.Such pick-up unit can bring owing to operation interpolation reaction reagent seems complicated for the operator, and can bring other influences to testing result simultaneously, because different additions can bring uncertain factor to testing result.These reactions are all finished in the liquid flow process in addition, in a single day sample is added in the pick-up unit, sample just goes ahead mobile, for example pass through sample region of acceptance 111 successively, marked region 121, surveyed area 132 usually makes reaction also not have the detection that fully just is not through with because of flowing of liquid.Utilize device of the present invention in advance the agent treated of needs and sample reaction on second reagent areas, the one, can allow sample and reagent reacting abundant, can not influence the degree that reaction is finished because liquid will in time flow, another is exactly the reagent that the operator does not need to add in addition other again.Like this, utilize pick-up unit of the present invention just can finish detection more simply fast, do not influence the accuracy of detection simultaneously.
Analyte can be can analyze any analyte with the present invention.The example of the analyte that the enough the present invention of energy detect comprises (but not only comprising) human chorionic gonadotrophin (hCG), lutropin (LH), ovarian stimulation element (FSH), hepatitis C virus (HCV), hepatitis B (HBV), hepatitis B surface antigen, AIDS virus.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, the bacterium composition is (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
In another concrete embodiment, utilize pick-up unit of the present invention to detect the small-molecule substance of sample analyte for end concentration, normally some haptens materials seem effective more and convenient.
In specific embodiment, based on competition law, comprise on second reagent areas with the incoherent specific bond molecule of analyte to it a kind of molecule M1, the molecule Y1 of this molecule and specific bond analyte links to each other or combination, and this reagent areas can be positioned at the upstream of first reagent areas on the reagent strip.More specifically, second reagent areas can be positioned at the upstream of the sample region of acceptance 11 of first reagent areas exactly, and sample region of acceptance 11 is positioned at the upstream of marked region 12, and marked region 12 is positioned at the upstream of surveyed area 13; On surveyed area, be fixed with the incoherent specific bond molecule of analyte to another kind of molecule M2, on marked region 12, comprise the similar substance A* (Fig. 5 A) of mark substance L and analyte.The first, the second with surveyed area on reagent concentration can be along with the different and any adjusting of the requirement that detects; For example in drugs detect, when need allow the concentration of certain drugs in the sample greater than the concentration C that sets in advance, allow the color lines on surveyed area, not occur and represent positive findings, in the time of less than concentration C, the color lines on surveyed area, occur and represent negative findings.
When reacting, if contain analyte A in the sample, and the time greater than the concentration C that sets in advance, then the specific bond molecule Y1 on second reagent areas almost completely carries out specific bond with analyte A and forms compound substance A-Y1-M1, may also have the excessive analyte A of residue this moment; When this compound substance arrives on the marked regions 12 by sample region of acceptance 11, because the reagent Y1-M1 on first reagent areas is by complete reaction, the reagent A * on the marked region-L just not can be incorporated on the Y1-M1; When this compound substance A-Y1-M1 flows through on the surveyed area 13, be fixed on the surveyed area with the incoherent specific bond molecule of analyte to another kind of this compound substance of molecule M2 specific bond, form M2-M1-Y1-A, thereby color showing positive findings (Fig. 5 B) on surveyed area, do not occur.
On the contrary, if contain analyte A in the sample, and less than the concentration C that sets in advance the time, then may also there be the excessive Y1-M1 of residue this moment in the carrying out specific bond with analyte A and form compound substance A-Y1-M1 of part just of the specific bond molecule Y1 on second reagent areas; When this compound substance arrives on the marked regions 12 by sample region of acceptance 11 because the reagent Y1-M1 on first reagent areas is also by complete reaction, the reagent A * on the marked region-L just and Y1-M1 in conjunction with formation M1-Y1-A*-L; As these this compound substances A-Y1-M1, when M1-Y1-A*-L flows through on the surveyed area 13, be fixed on the surveyed area with the incoherent specific bond molecule of analyte to another kind of this compound substance of molecule M2 specific bond, form M2-M1-Y1-A and M2-Y1-A*-L, thereby color showing negative findings (Fig. 5 B) on surveyed area, occurs.
These haptens materials comprise drugs (as drug abuse)." drug abuse " (DOA) is meant that the non-medical destination uses medicine (playing the paralysis nerve usually).Abuse these medicines and can cause body ﹠ mind to suffer damage, produce dependence, habit-forming and/or dead.The example of drug abuse comprises cocaine; Amphetamine (for example, black beauty, white amphetamine tablet, dextroamphetamine, dexie, Beans); Crystal methamphetamine (crank, meth, crystal, speed); Barbiturate (as Valium , Roche Pharmaceuticals, Nutley, NewJersey); Sedative (paramedicines of promptly sleeping); Lysergic acid diethylamide (LSD); Inhibitor (downers, goofballs, barbs, bluedevils, yellow jackets, methaqualone); The anti-antidepressant of tricyclic antidepressants (TCA, i.e. imipramine, amitriptyline and doxepin); Hog (PCP); Tetrahydrocannabinol (THC, pot, dope, hash, weed, etc.); Opiate (being morphine, opium, codeine, heroin, the hydroxyl dihydrocodeinone); Anxiolytic and hypnotic sedative agent, anxiolytic is that a class is mainly used in anxiety reduction, nervous, frightened, set the mind at rest, have the medicine of hypnosis sedation concurrently, comprise Benzodiazepines (benzodiazepines, BZ), atypia BZ class, merge phenodiazine NB23C class, the tall and erect class of benzene nitrogen, the part class of BZ acceptor, open loop BZ class, diphenylmethane derivatives, the piperazine carboxylic acid salt, the piperidine carboxylic acid salt, Kui azoles quinoline ketone, thiazine and thiazole, other heterocyclic, imidazole type calmness/anodyne, propanediol derivative-carbamates, fatty compound, anthracene derivative etc.Use this device also can be used to detect and belong to medical usage but the detection of overdose easily, as tricyclic antidepressant (imipramine or analog) and Paracetamol etc.Can resolve into different small-molecule substances after these medicines are absorbed by the body, these small-molecule substances are present in the body fluid such as blood, urine, saliva, sweat or there is above-mentioned small-molecule substance in part body fluid.
The similar substance of analyte is included in that above-mentioned analyte (haptens) go up to connect or coupling is associated with the antigenic substance that protein molecular can cause immune response, can also be the material of analyte other different chemical structures of deriving and the antigenic substance that is connected with immunogen protein.These haptens materials itself can not cause that immune response produces antibody, have only connection or coupling connection to go up immunogenic substances and could allow animal body produce antibody.These immunogenic substances comprise, but be not limited to this, albumen, nature or synthetic polypeptide or some carbohydrates, hemocyanin (Keyhole limpet hemocyanin for example, KLH), bovine hemoglobin (Bovine gamma globulin, BGG), bovine serum albumin (Bovine Serum Albumin, BSA), bovine thyroid albumen (Bovine Thyroglobulin, BTG), ovalbumin (Ovalbumin, OVA), sperm whale myoglobin (SpermWhale Myoglobin, SWM), tetanus toxoid (Tetanus Toxoid, TT), methylated bovine serum albumin (Methylated Bovine Serum Albumin, mBSA), human immunoglobulin(HIg) IgG or IgA (Humanimmunoglobulins IgG, IgA) or the immunogen protein of other prior aries.
These haptens are present in the different body fluid, detect with traditional pick-up unit, because the concentration of these small-molecule substances is sometimes very low, and in flow process, finish reaction, some micromolecule can be adsorbed by the reagent strip in the traditional detection device like this, particularly the sample region of acceptance is more important to the absorption of small-molecule substance, this suction-operated has reduced the amount of analyte relatively, thereby make the result of detection have deviation, sometimes or even wrong result with actual comparing.Utilize pick-up unit of the present invention at first allow in the sample analyte fully and the reagent on first reagent areas react, allow the sample that reacts completely flow to reagent strip more then and get on to finish detection reaction, can allow so at first in esse analyte and reagent carry out necessary complete reaction in the sample, after flowing on the reagent strip and then, these micromolecule analytes can not be attracted on the reagent strip, have reduced the interference of reaction.
More specifically in the examples of implementation, as Fig. 6, shown in 7,8, pick-up unit 30 comprises body 304 and collection well 601 at another, and wherein reagent strip 10 is arranged in body, and second reagent areas 201 is arranged in collection well 601; First reagent areas 201 and reagent strip 10 are in the liquid communication state.Here said " liquid communication state " is meant that liquid can flow on the reagent strip 10 from collecting chamber 601, this flowing can be owing to the action of gravity of liquid itself is flowed, also can flow through having through hole between collecting chamber 601 and the reagent strip 10, simultaneously between collecting chamber and reagent strip, can also install some flow guiding structures, can allow liquid flow on the reagent strip from collecting chamber more smoothly like this.More specifically, comprise on the body 304 and read window 3041 and sample ingress area 306, sample ingress area 306 comprises that sample imports through hole 306-1 and 306-3, read window 3041 correspondences in surveyed area on the reagent strip 10 and the body 304, the sample region of acceptance of reagent strip 10 and sample import through hole 306-1 correspondence (Fig. 8).Importing sample that through hole 306-1 flows into by sample can flow to sample region of acceptance on the reagent strip 10 and arrive surveyed area then and finish whole detection reaction.Collecting chamber 601 comprises first collecting chamber 302 and second collecting chamber 303, first collecting chamber, 302 1 end openings are used to admit the sample of needs detection or the gathering-device that admittance has sample, on the other end, have several liquid through-hole 302-1 that can allow liquid pass through, 302-2,302-3 (Fig. 7 A-D); One vertical bar beam 3024 is arranged on the correspondence position of liquid through-hole 302-1, and these beam 3,024 one ends can be fixed on the sidewall of first collecting chamber 302, and the other end extends downwards and passes liquid through-hole 302-1, and the second examination zone 201 is positioned on the vertical bar beam 3024.The second examination zone 201 can comprise reagent strip 2011, these reagent strip 2,011 one ends 2012 can be fixed on the surface of vertical beam 3024, the other end 2013 can pass the bottom that liquid through-hole 302-1 arrives second collecting chamber 303, on 2,013 one ends of reagent strip 2011, comprise can with the molecule (Y1) of the specific bond analyte of liquid flow and with the incoherent specific bond molecule of analyte to a kind of molecule (M1) in (M1/M2); The material of this reagent strip can be that some water-absorbing materials are formed, and can handle some protein substances in the above, for example glass fibre, filter paper, acetate fiber or the like, and an end 2012 of reagent strip 2011 can be bonded at the surface of vertical beam 3024 by organic nothing or machine glue.This reagent strip 2011 also can be adhesive among the liquid through-hole 320-3 or among the 302-2.First reagent areas 201 also can other form be present in the collecting chamber 601, for example reagent strip 2011 can be placed directly in the bottom of first collecting chamber 302, also can be placed in the insulating space that forms between first collecting chamber bottom and second collecting chamber bottom (Fig. 9), as long as reagent strip 2011 is placed in the collecting chamber 601, it is just passable to allow sample fully contact with it, 601 li of collecting chambers, reagent strip 2011 can be fully immersed in the sample, also can partly be immersed in the sample.In addition, the mode that second reagent areas exists also can be an alternate manner, can directly handle reagent in collecting chamber 601, and the mode of processing comprises sprinkling, smears or the like.Second reagent areas comprise a kind of can with the molecule (Y1) of the specific bond analyte of liquid flow and with the incoherent specific bond molecule of analyte to a kind of molecule (M1) in (M1/M2), in addition, can also comprise some other auxiliary reagents, the buffer reagent of buffer action for example, some protein moleculars or the like.Other forms or mode that one of ordinary skill in the art is all expected easily in conjunction with the present invention can be as one embodiment of the present of invention.First collecting chamber 302 and second collecting chamber 303 are assembled into collecting chamber 601 (Fig. 8), and collecting chamber 601 is connected as a whole (Fig. 8,9,10,11) by the snap ring 306-2 on the sample ingress area 306 on the body 304 with body 304.Second collecting chamber 303 comprises an end opening, and the other end has several liquid through-hole 303-3,303-2, and 303-1, at the corresponding O-ring seal 305-2 of being separately installed with of these liquid through-holes, 305-1 and 305-3 (Fig. 8, Fig. 9).Concretely, first collecting chamber 302 comprises an outer snap ring 3025, outer snap ring can directly block to scratch is having a structure to be connected with snap ring 306-2 on (Fig. 9) formation collecting chamber 601, the second collecting chamber lateral walls on the sidewall of second collecting chamber 303, and collecting chamber 601 can rotate in snap ring like this.Similar have concrete description with other concrete structrual descriptions this device in U.S. Patent application 2005/0180882A1.
Whether detect with collection saliva below exists drugs to come to describe how to use pick-up unit of the present invention in detail for example in the saliva.As shown in Figure 9, it is the collection phase before detecting, first collecting chamber 302 is positioned at second collecting chamber, 303 inside, and second the liquid through-hole 303-1 on the collecting chamber, 303-2,303-3 be not with sample ingress area 306 on sample import through hole 306-1,306-3 communicates, but sealed by the baseplane 306-4 of sample ingress area 3068, sealing for the baseplane 306-4 that increases collecting chamber 601 and sample ingress area 306, can O-ring seal 305-1,305-2 (omission), 305-3 be installed at the liquid through-hole correspondence position of the bottom of second collecting chamber 303.At first collect the saliva that to detect, collecting with sample collection device 301 earlier needs the saliva sample that detects, collect 3012 put into the detected person mouthful in, 3012 are made up of absorbent material, sponge material for example needs the saliva sample that detects so this collection head can absorb.Then suction is had in collection 3,012 first collecting chamber of putting in the collecting chamber 601 302 of saliva sample, head is collected in extruding, and at this time the saliva sample of collecting on the head just is extruded in first collecting chamber 302, and by liquid through-hole 302-1,302-2,302-3 flow in two collecting chambers 303.In this process, because first reagent strip 2011 in first collecting chamber extends to the bottom of second collecting chamber 303 by liquid through-hole, in the extruding sample, saliva sample just and the reagent on first reagent strip 2011 fully react, after reacting suitable time by the time, as 10,20 or 30 seconds, 1,2,3,4,5,6,7,8,9,10,15,20,30,45, after 60 minutes, rotate collecting chamber 601, allow the liquid through-hole 303-1 and the importing of the liquid on the liquid ingress area through hole 306-1 of second collecting chamber communicate, liquid through-hole 303-3 and liquid import through hole 306-3 and communicate.This time, saliva sample just flows to by liquid importing through hole 306-1 and carries out detection reaction on the reagent strip 10, if there are a certain amount of drugs in the saliva, the color lines on the surveyed area of reagent strip, do not occur and represent positive findings, just occur the color lines on the contrary and represent negative findings, this testing result on surveyed area can be by reading window 3041 observationss on the body, and reading on the window 3041 can be that material transparent covers surveyed area and also can directly allow surveyed area exposed outside.Saliva sample can also import through hole 306-3 by liquid and enter and confirm chamber 307, detects saliva sample in order further to confirm.Confirm that chamber 307 is surrounded by base 3071 and sidewall on every side, the liquid that enters the affirmation chamber can take out by deriving through hole 3073, derives through hole 3073 and is sealed by a stopper 3072 before sampling.When thinking that testing result needs further by other means, when for example with the liquid phase look general, gas chromatography or other more accurate instruments are confirmed testing result, remove stopper 3073, take out the part saliva sample and detect (as Fig. 8,10 and 11) from confirming chamber 307.Here only being to describe how to use pick-up unit of the present invention with saliva sample in detail as an example, except saliva, can also be other liquid samples, for example blood, urine, sweat, ight soil or the like.
In another concrete embodiment, second reagent areas not necessarily will with reagent strip in same pick-up unit, it can also with gathering-device that pick-up unit separates in or in other containers.Concretely, on second reagent areas, comprise exactly a kind of specific bond analyte molecule and with the incoherent specific bond molecule of analyte to it a kind of molecule, surveyed area on reagent strip comprise fixing and the incoherent specific bond molecule of analyte to another kind of molecule, wherein second reagent areas is arranged in gathering-device or other containers that separates with pick-up unit.For example, second reagent areas can be arranged in a test tube, and the reagent that sample is joined on second reagent areas in test tube relief sample and the test tube reacts, and the reaction liquid in the test tube is joined finish whole detection on the reagent strip then.These gathering-devices and container can be listed in U.S. Pat 6780160 down; US7048693; US5234001; US5830154; US5786427; Find among the US5573099, can also in application is open, find, for example US2001/0008614; WO2005008216 or the like.
Experiment is now given description of test for more detailed elaboration pick-up unit of the present invention.
Experiment 1: detect in the saliva whether have certain density anxiolytic and hypnotic sedative agent (BZO) with pick-up unit
Anxiolytic (being called for short BZO) is that a class is mainly used in anxiety reduction, anxiety, fear, set the mind at rest, have the medicine of hypnosis sedation concurrently, comprise Benzodiazepines (benzodiazepines, BZ), part class, open loop BZ class, diphenylmethane derivatives, piperazine carboxylic acid salt, piperidine carboxylic acid salt, Kui azoles quinoline ketone, thiazine and the thiazole of atypia BZ class, the phenodiazine NB23C class that merges, the tall and erect class of benzene nitrogen, BZ acceptor, other heterocyclic, imidazole type calmness/anodyne, propanediol derivative-carbamates, fatty compound, anthracene derivative etc.These drug abuses also can produce dependence and be harmful.
First: the assembling of reagent strip of the present invention and pick-up unit and parts are made (one of the present invention's experiment)
The processing of nitrocellulose filter.Detection lug 131 is nitrocellulose filter (NC), and two lines are arranged on nitrocellulose filter, and one is detection line 207, is positioned at the testing result control line 206 in detection line downstream.On detection line, be fixedly connected with the Streptavidin (streptavidin-IgG) of IgG; Fixing foster anti-rabbit igg on the testing result control line.Fixing method can be handled automatically with spraying the film handling machine automatically, is 0.3 mg/ml handling the concentration that detects lines wherein, and dilution buffer liquid is phosphate buffer (PBS); Fixing goat anti-rabbit igg antibody on the testing result control line; Concentration is 0.3 mg/ml, and dilution buffer liquid is phosphate buffer (PBS).The nitrocellulose filter of handling well is placed on to dry in 37 ℃ of baking ovens gets final product.
The processing of marker slip 121.Marker slip is the polyester film, and the have coupled protein good by the colloid gold particle mark divides the BZO haptens of BSA to be processed on the polyester film.The OD value of Treatment Solution is 75, and dilute solution is 1 times of PBS buffer solution, wherein also contains 1% BSA.On marker slip, also handle the rabbit igg antibody that colloid gold label is arranged.The marker slip of handling well is placed on to dry in 37 ℃ of baking ovens gets final product.
The processing of sample blank film 111.The sample blank film is the plain film of glass fibre, and the solution of handling on this film is: Borax (0.07M/L); Tween20 (1%); Cholic Acid (1%); Tris (0.1M).The sample blank film of handling well is placed on to dry in 37 ℃ of baking ovens gets final product.
The assembling of reagent strip.According to appearance assembling shown in Figure 2, allow each parts of handling well the sample blank film be positioned at the upstream of marker slip, marker slip is placed the filter paper of suction in the downstream of detection lug between detection lug and sample blank film.These parts all are placed on the non-absorptive support chip.
The processing of second reagent areas 201.Second reagent areas comprises a reagent strip 2011, and this reagent strip 2011 comprises non-absorptive paste-side 2012 and absorptive reagent end 2013.The material of reagent end is a polyester film, and the chemical solution that the reagent end is handled comprises: be dissolved in 1 times of PBS buffer solution connecting the anti-BZO antibody materials of going up biotin (Biotin), making ultimate density is 0.15 mg/ml; Wherein also contain 1% BSA.The reagent strip of handling well 2011 is placed on to dry in 37 ℃ of baking ovens gets final product.Then the polyester film small pieces (2013) that contain reagent are sticked on the non-absorptive small pieces on (2012).
The assembling of pick-up unit.Pick-up unit is gone into Fig. 6, shown in 7,9.The paste-side 2012 of reagent strip 2011 is sticked on the surface of the vertical bar beam 3024 on first collecting chamber 302, the other end 2013 passes liquid through-hole 302-1 and arrives the bottom of second collecting chamber 303.Then whole collecting chamber 601 is installed among the snap ring 306-2 of sample Lead-In Area 306 of body 304, and allows the bottom of second collecting chamber 303 be in the position of the basal surface 306-4 sealing (Fig. 9) that is imported into the zone.Reagent strip 101 is placed between upper plate 3043 and the lower plate 3045, makes the surveyed area of reagent strip 101 corresponding with reading window 3041, sample region of acceptance and liquid import through hole 306-1 correspondence.It is an integral body that upper plate and lower plate fasten.So just be assembled into the used pick-up unit of this experiment.
Second portion: the assembling of control stripes bar and pick-up unit and making (common competition law detects drugs).
Compare with pick-up unit with reagent strip recited above, the antibody of handling on the contrast agents bar label pad by the good anti-BZO of colloid gold particle mark is processed on the polyester film.With 1 times of PBS buffer solution configuration flag solution, the OD value of ultimate density is 75, wherein also contains 1% BSA.On marker slip, also handle the rabbit igg antibody that colloid gold label is arranged.The marker slip of handling well is placed on to dry in 37 ℃ of baking ovens gets final product.
Fixedly having the BZO haptens that coupled protein divides BSA on the detection line that detects on the film, is 0.3 mg/ml handling the concentration that detects lines wherein, and dilution buffer liquid is phosphate buffer (PBS).The reagent that other parts of contrast agents bar are handled is the same with the description of first with condition.
In order to check superiority of the present invention, do check with same sample.This sample is a saliva sample, and the concentration that BZO is set respectively in saliva sample is 5ng/ml (nanograms/milliliter); 7.5ng/ml; (detection threshold, cut off) 10ng/ml; 12.5ng/ml; 15ng/ml; 30ng/ml.Testing result record all reads after 10 minutes adding saliva sample.The meaning of detection threshold just be meant if the concentration of the analyte BZO in the sample greater than detection threshold, in theory, testing result should be positive, otherwise, if the concentration of analyte BZO less than detection threshold, testing result is negative.
Method of operating:
Allow each pick-up unit be in position shown in Figure 9 earlier, promptly the bottom of second collecting chamber 303 of collecting chamber 601 is imported into the 306-4 sealing in zone.
Add the sample that to detect then, allow in sample and the collecting chamber 601 reagent reacting on second reagent areas 1 minute;
The actual result that record detects after 10 minutes.
Experimental result:
Control test device testing result and analysis
| The BZO sample | Handle | Experimental result | Correct testing result | Recall rate | |||||||||||
| Negative | Positive | ||||||||||||||
| Lot #1 | Lot #2 | Lot #3 | Lot #1 | Lot #2 | Lot #3 | Lot #1 | Lot #2 | Lot #3 | Lot #1 | Lot #2 | Lot #3 | Lot#1 | Lot#2 | Lot#3 | |
| |
10 | 10 | 10 | 10 | 10 | 10 | 0 | 0 | 0 | 10 | 10 | 10 | 100％ | 100％ | 100％ |
| 5ng/ |
10 | 10 | 10 | 10 | 10 | 10 | 0 | 0 | 0 | 10 | 10 | 10 | 100％ | 100％ | 100％ |
| 7.5ng/ |
10 | 10 | 10 | 10 | 10 | 10 | 0 | 0 | 0 | 10 | 10 | 10 | 100％ | 100％ | 100％ |
| 10ng/ml (Cut off) | 10 | 10 | 10 | 9 | 10 | 10 | 1 | 0 | 0 | N/A | N/A | N/A | N/A | N/A | N/A |
| 12.5ng/ |
10 | 10 | 10 | 9 | 9 | 10 | 1 | 1 | 0 | 1 | 1 | 0 | 10％ | 10％ | 0 |
| 15ng/ |
10 | 10 | 10 | 8 | 9 | 10 | 2 | 1 | 0 | 2 | 1 | 0 | 20％ | 10％ | 0 |
| 30ng/ |
10 | 10 | 10 | 8 | 9 | 9 | 2 | 1 | 1 | 2 | 1 | 1 | 20％ | 10％ | 10％ |
Laboratory test results of the present invention and molecule result:
| The BZO sample | Handle | Experimental result | Correct testing result | Recall rate | |||||||||||
| Negative | Positive | ||||||||||||||
| Lot #1 | Lot #2 | Lot #3 | Lot #1 | Lot #2 | Lot #3 | Lot #1 | Lot #2 | Lot #3 | Lot #1 | Lot #2 | Lot #3 | Lot#1 | Lot#2 | Lot#3 | |
| |
10 | 10 | 10 | 10 | 10 | 10 | 0 | 0 | 0 | 10 | 10 | 10 | 100％ | 100％ | 100％ |
| 5ng/ |
10 | 10 | 10 | 10 | 10 | 10 | 0 | 0 | 0 | 10 | 10 | 10 | 100％ | 100％ | 100％ |
| 75ng/ |
10 | 10 | 10 | 10 | 10 | 10 | 0 | 0 | 0 | 10 | 10 | 10 | 100％ | 100％ | 100％ |
| 10ng/ml (Cut off) | 10 | 10 | 10 | 5 | 5 | 6 | 5 | 5 | 4 | N/A | N/A | N/A | N/A | N/A | N/A |
| 12.5ng/ |
10 | 10 | 10 | 3 | 3 | 4 | 7 | 7 | 6 | 7 | 7 | 6 | 70％ | 70％ | 60％ |
| 15ng/ |
10 | 10 | 10 | 2 | 2 | 2 | 8 | 8 | 8 | 8 | 8 | 8 | 80％ | 80％ | 80％ |
| 30ng/ |
10 | 10 | 10 | 0 | 1 | 0 | 10 | 9 | 10 | 10 | 9 | 10 | 100％ | 90％ | 100％ |
Result and conclusion:
Can clearly find out from result of upper experiment, utilize the analyte that detects low concentration in the sample that device of the present invention can be very sensitive, greatly the sensitivity of the detection of Ti Gaoing and do not change specificity.Utilize competition law to detect BZO in the saliva in the prior art, when the concentration of BZO in the sample was 12.5, having only 2, can to detect be positive, all the other 28 complete negative results, and recall rate is about 10%; Yet, utilize pick-up unit of the present invention that 20 positive results are arranged, 10 negative findingses, recall rate is 60%-70%; Equally, when the concentration of BZO in the sample was 12.5 and 15, the recall rate of the pick-up unit of prior art was about 10%-20%; And the recall rate of utilizing pick-up unit of the present invention to reach is 80%-100%, is far longer than the pick-up unit of prior art.In addition, in 30 negative sample, pick-up unit of the present invention is all negative, and the specificity that does not change pick-up unit is described, promptly is not subjected to the interference of negative sample.
Claims (18)
1. whether one kind detected and exist the pick-up unit of being analysed material to comprise in the sample: reagent strip, on this reagent strip, comprise first reagent areas and surveyed area, wherein, described surveyed area comprises a kind of fixed specific bond molecule, first reagent areas is positioned at the upstream of surveyed area, it is characterized in that: described pick-up unit also comprises second reagent areas of separating with described reagent strip, this second reagent areas comprise a kind of can be with the molecule of the specific bond analyte of liquid flow; Described second reagent areas and described reagent strip are in the liquid communication state.
2. pick-up unit as claimed in claim 1 is characterized in that: described second reagent areas also comprise with the incoherent specific bond molecule of analyte to it a kind of molecule; Described fixed specific bond molecule comprise with the incoherent specific bond molecule of analyte to another kind of molecule.
3. pick-up unit as claimed in claim 2, described first reagent areas comprises the sample region of acceptance and contains the marked region of mark substance, the sample region of acceptance is positioned at the upstream of marked region, and described second reagent areas is positioned at the upstream of sample region of acceptance and is in the liquid communication state with the sample region of acceptance.
4. pick-up unit as claimed in claim 3, the molecule of the specific bond analyte on described second reagent areas comprises the first antibody of analyte, described marked region also comprises the second antibody of analyte.
5. pick-up unit as claimed in claim 3, the molecule of the specific bond analyte on described second reagent areas comprises the antibody of analyte, described marked region also comprises the similar substance of analyte.
6. pick-up unit as claimed in claim 5, analyte comprise drugs class material, and marked region comprises colored mark substance, and this colored mark substance is selected from one of colloid gold label particle, latex marking particle or water-soluble mark substance.
7. pick-up unit as claimed in claim 2, described and the incoherent specific bond molecule of analyte to be selected from following molecule to one of: biotin/affinity element (biotin/avidin); Biotin/Streptavidin (biotin/streptavidin); The antibody of rhodamine/rhodamine (rhodamine/anti-rhodamine); The antibody of mouse IgG/ mouse IgG (Mouse IgG/anti-mouse IgG).
8. pick-up unit as claimed in claim 1, described surveyed area comprise zone and the testing result control area that shows testing result, and this control zone is positioned at the downstream of surveyed area, also comprise suction zone in the downstream of testing result control zone; Wherein, the zone of demonstration testing result comprises described a kind of fixed specific bond molecule.
9. pick-up unit as claimed in claim 1, the surveyed area of described reagent strip is positioned on the nitrocellulose filter.
10. whether there is the method for analyte in the tracer liquid sample, comprises:
A kind of pick-up unit is provided, this pick-up unit comprises the reagent strip and second reagent areas, described reagent strip comprises first reagent areas and surveyed area, and wherein first reagent areas comprises sample region of acceptance and marked region, and surveyed area comprises a kind of fixed specific bond molecule; Described second reagent areas comprise a kind of can be with the molecule of the specific bond analyte of liquid flow, and described second reagent areas is positioned at the upstream of sample region of acceptance on the reagent strip, and and described reagent strip be in the liquid communication state; It is characterized in that:
Annex solution sample body on second reagent areas, allow this liquid sample earlier with second reagent areas on reagent reacting after 30 seconds-60 minutes;
Allow liquid sample flow to then and finish whole detection on the described reagent strip;
Read testing result.
11. method as claimed in claim 10, it is characterized in that: on the described surveyed area fixed specific bond molecule comprise with the incoherent specific bond molecule of analyte to it a kind of molecule, described second reagent areas comprise with the incoherent specific bond molecule of analyte to another kind of molecule, the molecule of this molecule and described specific bond analyte link to each other or in conjunction with and can flow with liquid.
12. method as claimed in claim 11, it is characterized in that: also comprise marked region on the described reagent strip, marked region comprises mark substance, this marked region is positioned at the downstream of sample region of acceptance, and this marked region is positioned at the upstream of surveyed area, and wherein the mark substance on the marked region links to each other or combination with the analyte analog.
13. method as claimed in claim 12 is characterized in that: behind the second reagent areas annex solution sample body, allow reagent reacting 30 second-30 minute on the liquid sample and second reagent areas;
14. method as claimed in claim 13 is characterized in that: allow on the liquid sample and second reagent areas reagent reacting 1-30 minute.
15. method as claimed in claim 14 is characterized in that: described liquid sample is one of saliva, urine or blood; Described analyte is a drugs micromolecular material.
16. method as claimed in claim 15 is characterized in that: described liquid sample is a saliva; Described analyte is THC or BZO class material.
17. one kind is detected the kit that whether has analyte in the sample, comprising: pick-up unit and gathering-device,
Wherein, pick-up unit comprises reagent strip, described reagent strip comprises the sample region of acceptance, marked region, surveyed area and suction zone, the sample region of acceptance is positioned at the upstream of marked region, marked region is positioned at the upstream of surveyed area, surveyed area is positioned at the upstream of suction zone, comprises that on surveyed area a kind of fixed and the incoherent specific bond molecule of analyte are to it a kind of molecule
Wherein, gathering-device comprise can the absorption liquid sample body the collection head;
It is characterized in that, in described pick-up unit, also comprise second reagent areas of separating with reagent strip, wherein, this second reagent areas comprise with the incoherent specific bond molecule of analyte to another kind of molecule and a kind of molecule of specific bond analyte.
18. kit as claimed in claim 17, described marked region comprises the similar substance of colored mark substance and analyte.
Priority Applications (9)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2006100526281A CN101017169B (en) | 2006-07-26 | 2006-07-26 | Analysis equipment of biological sample |
| CA002658795A CA2658795A1 (en) | 2006-07-26 | 2007-07-24 | A test device for detecting an analyte in a liquid sample |
| PCT/CN2007/070344 WO2008014709A1 (en) | 2006-07-26 | 2007-07-24 | Analysis device for biologicla sample |
| AU2007280929A AU2007280929B2 (en) | 2006-07-26 | 2007-07-24 | Analysis device for biological sample |
| DE212007000054U DE212007000054U1 (en) | 2006-07-26 | 2007-07-24 | Test device for detecting an analyte in a liquid sample |
| NZ575068A NZ575068A (en) | 2006-07-26 | 2007-07-24 | Analysis device for determining the presence of an analyte comprising a test strip having a separable second reagent zone. |
| JP2009600025U JP3157356U (en) | 2006-07-26 | 2007-07-24 | Analytical equipment for biological samples |
| US12/360,087 US8071394B2 (en) | 2006-07-26 | 2009-01-26 | Test device for detecting an analyte in a liquid sample |
| CN201010005320.8A CN101893626B (en) | 2006-07-26 | 2010-01-15 | Test device for detecting analyte in liquid sample |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
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