CN1125984C

CN1125984C - Assay device

Info

Publication number: CN1125984C
Application number: CN94194176A
Authority: CN
Inventors: 劳夫·A·古伊尔古斯
Original assignee: RAMINA CO Ltd
Current assignee: RAMINA CO Ltd
Priority date: 1993-09-17
Filing date: 1994-09-16
Publication date: 2003-10-29
Anticipated expiration: 2014-09-16
Also published as: JPH09512333A; BR9407504A; CN1135259A; AU7797994A; FI961183A0; EP0720744A1; WO1995008117A1; KR100314101B1; EP0720744A4; CA2171968A1; FI961183A; JP3529096B2

Abstract

An assay device and method are provided which allow for determination of the presence of an analyte in a sample while providing identification of the test subject. Self-contained assay device (110) includes a housing (111) with a first part (111a) and a second part (111b). Reaction medium (114) is located within the first part (111a) and mounted on a base member (112). Base member (112) includes a porous support which may be adjacent to the reaction medium (114) which includes at least reaction zone (120) as well as a control zone (116). A medium (126) containing a signal-producing agent is located in the second part of the housing (111b) and mounted on a base portion (128). The device additionally comprises a cylindrical body (140) having an open end (140a) and a closed end (140b) on which is mounted a medium (185) for containing and transferring a portion of test sample to the reaction medium (114).

Description

A kind of mensuration verifying attachment and method of providing for oneself

The application is to be on September 14th, 1992 applying date, application number is the part continuation application of the international application of PCT/US92/07785, and be to be on September 9th, 1993 applying date but not receive the part continuation application of the continuation application of application number as yet, this application of not receiving series number as yet is 07/759 at the application number of the U.S., 922, the applying date is on September 13rd, 1991 (this application in the U.S. is PCT/US 92/07785 a desired priority application), should be to be on March 12nd, 1991 applying date in the application of the U.S., application number is 07/668, the part continuation application of 115 U. S. application (now abandoning), this resigned U. S. application is that application number is 07/467,532, the applying date is the part continuation application of the U.S. Patent application (this application is now abandoned) in January 19 nineteen ninety, introduces all these applications as reference here.

TECHNICAL FIELD OF THE INVENTION

The invention belongs to ligand-receptor determination field, this mensuration comprises immunoassays, is used for determining that a kind of biofluid has or not certain analyte.More particularly, apparatus and method of the present invention relate to the identity that set up to confirm checked object and determine to take from an independent verifying attachment in certain biofluid of this checked object and have or not at least a analyte.

Background of invention

For many years, the those of ordinary skill in ligand-receptor determination field is being sought a kind of effective, cheap and reliable apparatus and method always, and being used for detecting has and/or do not have antigen, antibody and analog.This field has been full of such device.

For many years, those of ordinary skill in the art also seeks always and is used to guarantee the sample reality that the is verified apparatus and method from a certain definite individuality.Established many schemes, some is complicated, and some is simple, is in fact produced by a specific individuality so that confirm check sample.Similarly, having many schemes to hinder makes a certain specific assay consistent with a certain specific people.

For example, now drug abuse test is a kind of conventional program for sportsman, prisoner and in the workplace.Those records relate to the single fluid sample of check, for example urinate or blood, so that determine to exist in this fluid sample some specific antibody; Positive result can show and used medicine.

Before, finish such check by a series of tests, this campaign relates to a fluid that is verified and moves on in some different containers, and this fluid is got at a distance from the person that is verified.Often misplace and/or replace the problem of fluid with regard to the fluid that proposed relevant a series of supervision and be verified.Like this, a problem check fluid that occurs in such inspection period may be to obtain from some other people there, rather than that people's acquisition from being verified, perhaps, after the check fluid returned from testing laboratory, those check fluids can mix, lose or can not specifically be identified was for which people.Also have, these are measured generally needs the long time, so that can not in time obtain the result.

The many apparatus and method that are used for the analyte of definite biofluid at present also have some other shortcomings.Often need technician's use of certain level or handle such device, so that obtain satisfied result.In addition, the overwhelming majority of these devices needs one or more other reagent, and often needs rinse solution, so that be applied to during the checkout procedure on this device.For many such devices, to small part be because the limited susceptibility of device, need than relatively large, be expensive reagent or often than relatively large check sample, so that provide accurate result.

As mentioned above, the principle that constitutes the basis of this type checking is individual's a immune system, i.e. mammal response external, generally be the capability of the molecule of macro molecules.After this, any molecule that can draw a kind of like this response will be called antigen, and promptly antibody produces the source.The protein and the protein fragment that produce when response antigen will be called antibody or immunoglobulin (Ig).

Summary of the invention

The present invention by adopt custom-designed determinator overcome in known those device and the system intrinsic those problems, preferably, described custom-designed determinator is the feasible a kind of immunoassay apparatus that can determine to have or not in the sample certain analyte, and preferably, the special discriminating of this immunoassay apparatus provides the people of described sample.

The invention relates to the method for inspection and device, and more particularly, is to relate to specially being used to detect the method and apparatus that certain analyte exists, and this analyte is some specific antigens or the specific antibodies in certain biofluid most preferably.The present invention also preferably is used for differentiating reliably the individual who is verified.

The invention provides a kind of easy operating, the disposable test pad, and a kind of disposable test device, this verifying attachment are suitable for detecting and have or not certain analyte, and preferably suitable for the diagnostic test object.For example, available one ligand that has mark applies on the finger of checked object, and this finger is pressed on a kind of film substrate with one specific, ligand layer of having fixed, so that from biofluid, catch certain analyte, and provide (specific combination), the visible impression of the hand of the no prepared Chinese ink of checked object simultaneously, thereby reach indisputable ground, differentiate the supplier of described fluid reliably.Resulting fluid supplier's impression of the hand also can comprise the information about some analytes, for example medicine that exists in his body fluid when testing on film.This information can write down or store, and when needs, is used for differentiating reliably.

An object of the present invention is to measure a check sample and have or not certain analyte, and also differentiate the object that is verified simultaneously.Like this, can eliminate mistake evaluation, a series of icon and delay issue.

An object of the present invention is to provide the device whether a kind of certain analyte that is used for determining fast taking from a checked object exists.A further object of the invention provides a kind of device of amount that certain analyte of sample of a checked object is determined to take from fast quantification ground that is used for.And another purpose of the present invention provides a kind of apparatus and method of determining to have or not certain medicine or drug metabolite in the check sample of taking from a checked object, and provides an a kind of definite checked object whether to abuse certain device acute or chronic medicine.

Be not crucial with impression of the hand diagnostic test object in an alternative embodiment of the invention, this embodiment relates to the ligand of humoral sample or tape label is applied on the film as the transfer device of the part of verifying attachment or the pressure of spreader with one.With the surface that liquid check sample and/or some reagent drop by drop is applied to a kind of reaction medium, similarly, those known devices compare, in the present invention, liquid check sample and/or some reagent are applied on the reaction medium surface under pressure, and the time that keeps one section weak point on this reaction medium surface, thereby the present invention needs the check sample and the reagent of less amount and has shown higher susceptibility than those known devices.

Adopting the device of this principle according to the present invention is a kind of mensuration verifying attachment of sealing.In a preferred embodiment, this mensuration verifying attachment comprises a shell, a kind ofly has at least one to be arranged in the reaction medium of the conversion zone of this shell, a kind of a kind of reagent signal or that show bright analyte and a kind of medium that is used for holding a kind of check sample that is positioned at above-mentioned shell of producing that is arranged in this shell that contain.Contain the medium of the reagent that produces signal and be used for holding the medium both of a check sample of above-mentioned shell and can carry out independent moving between the second place that transferable material contacts at a primary importance that separates each other with above-mentioned reaction medium and one and described reaction medium.Preferably, this is a kind of verifying attachment of providing for oneself.

A certain embodiments of the present invention comprises a mensuration verifying attachment that has adopted the shell of one two parts.In an embodiment of this device, a kind of reaction medium with at least one conversion zone is installed in the first of described shell.Be arranged in above-mentioned two parts shell second portion be a kind of medium that is used to hold a part of check sample.Be used to hold among the medium of certain check sample of a part and two parts that reaction medium is arranged in above-mentioned shell described like this, make when each case member in the sealing mode and when lumping together, have the contact of transferable material between described medium that is used to hold a part of certain check sample and the reaction medium.Except at least one conversion zone, above-mentioned reaction medium preferably has one to check the zone, disposes this and checks the zone so that come the diagnostic test object by impression of the hand or similar means, if be ready like this.So a kind of device can be designed to a kind of determinator of providing for oneself.Used herein, term " is provided for oneself " and is meant and can uses this device and need not additional other reagent or solution, perhaps, just need drop by drop add water.

Another embodiment has adopted some similar device, but has replaced being used to hold the medium of certain check sample of a part with a kind of medium that contains the reagent that produces signal.

A kind of preferred device according to above-mentioned " pressure " principle is similar to aforementioned those devices, similar part has been also to adopt the shell of one two parts, in this shell, the reaction medium with at least one conversion zone is installed in the first in the shell of these two parts.But,, in the second portion of the shell of described two parts, be provided with a kind of medium that certain produces the reagent of signal that contains, for example, show the material that has mark that bright certain analyte exists as one of these embodiment.Described device also comprises a kind of removable medium that is used to hold certain check sample of a part, this kind check sample is positioned at the centre of first and second parts of above-mentioned shell, and when first and second parts of described shell are positioned at one first detent position of this shell in the sealing mode, carry out contacting of transferable material with described reaction medium.Locate and dispose reaction medium like this and contain certain reagent that shows bright analyte (or reagent of generation signal) and be among their case members separately, make that described reaction medium is in contacting of transferable material each other with the device that is used for shifting the reagent that shows bright analyte when each parts of described shell are in a second place in the sealing mode.

When using this device, the check sample that extracts from a checked object is placed on a kind of removable medium that is used to hold and shift a part of check sample, and this check sample is placed among the shell of two parts.Then, this shell is closed to a primary importance, thereby the removable check sample that contains medium is in the contacting of transferable material with reaction medium and exerts pressure for this reaction medium.Then, open described shell, and from this shell, remove the removable medium that is used to hold and shift certain check sample.Make described shell be closed to one second detent position for the second time, be in above-mentioned reaction medium is transferable and contact and exert pressure for this reaction medium thereby be used for shifting certain device that shows the reagent of bright analyte.

In another embodiment of the present invention, the various segregate zone of reaction medium comprises the specific composition of a kind of immunity to (MIP), and this immunity is to combination or catch certain specific analyte, for example, and certain medicine or drug metabolite.Different segregate zones can comprise some MIP that are specifically designed to some appointed different analytes, and thus, making can be to determining more than a kind of analyte in once independent mensuration.

Method and apparatus advantageous particularly part of the present invention only need with traditional moving phase ratio that passes through some boxes or device to be the reagent of a spot of generation signal; When keeping directly contact, bring pressure to bear on the local concentration that has kept some reagent on the surface of verifying attachment, particularly produce the reagent of signal, this reagent presents increases reaction velocity and susceptibility, for example, about second resulting assay with about divide or resulting assay of longer time relatively; And in certain embodiments, mark secondary antibodies (opposite with mark primary antibody or antigen) reduces the reaction time of measuring, and this is because by antigen being attached to the shape that does not change this antigen on the mark.

Definition

1. ligand-receptor determination: any technology that relates to the detection that is formed on the compound between a kind of ligand and a kind of material that is attached on this ligand.Preferably, a kind of composition of this compound is a kind of analyte.Preferred ligand-receptor determination is a kind of immunoassay.Ligand-receptor determination can be used to determine the having of in biofluid ligand, nothing, quantity and concentration.Ligand-receptor determination can be emulative or noncompetitive technology, single or complicated technology, direct or indirect technology or a kind of combine detection technology, for example, a kind of antibodies to a small half of chemicals, biological example element or dinitrophenol (DNP).

Preferably during the mensuration process, all basically at least a predetermined ligand or ligand acceptors are retained in a precalculated position.Any technology that is used for fixing a kind of ligand or ligand acceptor all comprises within the scope of the invention.In a preferred embodiment, a kind of ligand or ligand receptors bind or be fixed on that a kind of solid is gone up mutually or wherein.Typical fixed mechanism includes, but are not limited to covalent bond, non-covalent combination, chemical coupling, physics captures and absorption.

2. ligand: be meant analyte itself or can be used for inferring a kind of material that in a check sample, has certain analyte to exist.A kind of ligand-acceptor is meant some materials, and these materials are had a specific binding partner, for example, and ligand.Used herein, ligand and ligand-acceptor can be the member of an immunity to (MIP).Ligand and ligand acceptor can comprise the metabolin of haptens, hormone, antigen, antibody (comprise anti--antibody and antibody fragments, for example, Fab or Fc), DNA, RNA, oligonucleotide, nucleic acid and some compounds or above-mentioned any material.Ligand or ligand-acceptor can tape labels or tape label not.

3. analyte: determined material, and, depend on used specific mensuration, can be a kind of ligand or a kind of ligand-acceptor.Some typical analytes include, but are not limited to the metabolin of medicine, protein, haptens, hormone, aforementioned these materials, and some other molecules, independent or with a kind of protein bound composition together.A kind of haptens is a kind of small molecular weight material, for example some drugs or drug metabolite, haptens generally at first is attached on a kind of protein, for become antigen or immunity.For example, can find the medicine of some freedom and combining form in vivo, this point is transferred to make and can be distinguished chronic drug abuser and acute drug misuser.For the former situation, in many examples, form certain antibody of opposing combining form in vivo, the also available the present invention of this antibody detects and is a kind of analyte.

4. biofluid: can be verified so that determine to have or not any body fluid of certain analyte, include, but are not limited to blood, blood component, saliva, urine.

5. produce the reagent of signal: be used in a kind of ligand-receptor determination, can be used to produce any reagent of a detectable signal.Some reagent that preferably produce signal provide a kind of signal that is easy to estimate, and more preferably a kind of color.

The accompanying drawing summary

Fig. 1 is a planimetric map of a kind of determinator of the present invention;

Fig. 2 is a planimetric map of a kind of determinator of the present invention, and it shows the result who has or not certain antigen check and is suitable for differentiating the formation of the pattern of a checked object;

Fig. 3 is the schematic cross sectional view that of a kind of determinator of the present invention has amplified, and it demonstrates a conversion zone in a kind of reaction medium;

Fig. 4 is the schematic cross sectional view that of a kind of determinator of the present invention has amplified, and its of demonstrating in a kind of reaction medium checks the zone;

Fig. 5 is the schematic cross sectional view that of a kind of determinator of the present invention has amplified, and it demonstrates the assay of a positive;

Fig. 6 is the schematic cross sectional view that of a kind of determinator of the present invention has amplified; It demonstrates the assay of a feminine gender;

Fig. 7 is a recapitulative view of a kind of determinator of the present invention, and it demonstrates the contact of determinator of the present invention and method;

Fig. 8 is a recapitulative sectional view of a kind of immunoassay apparatus of the present invention, and it demonstrates a formation of checking an impression of the hand in the zone at reaction medium;

Fig. 9 is the recapitulative sectional view of another embodiment of a kind of immunoassay apparatus of the present invention, and it demonstrates the transfer on the finger of a kind of test mixture from swab to the people that check sample is provided;

Figure 10 is a recapitulative sectional view of the immunoassay apparatus of Fig. 9, and it demonstrates by the above-mentioned people's of check sample the finger that provides described test mixture is applied on the described reaction medium;

Figure 11 is open, the expansion skeleton view of an embodiment of a kind of device of the present invention;

Figure 12 is the skeleton view of another embodiment of a kind of device of the present invention;

Figure 13 is the skeleton view of another embodiment of the present invention.

Detailed description of the present invention

Ligand receptor determination device of the present invention comprises a kind of reaction medium, and this reaction medium has a conversion zone at least, and preferably, has a zone of checking that can confirm a checked object identity at least.In a preferred embodiment of the present invention, described conversion zone comprises a kind of ligand, ligand-acceptor at least to the member of (also being called a kind of " ligand/acceptor to " herein), and this member can confirm to have or not certain material in a check sample.In another preferred embodiment of the present invention, be incorporated into a kind of ligand in the zone so that provide the authentication pattern of a checked object to differentiate the identity of above-mentioned checked object by checking at least one.

Method of the present invention be included in a part certain reaction medium on or wherein, carry out in a check sample, having or not the immunoassays of certain analyte, and confirm on the described reaction medium of a part, to provide the identity of the checked object of above-mentioned check sample independently.

One embodiment of the present of invention are at a kind of determinator, preferably at a kind of immunoassay apparatus that comprises the reaction medium of a porous bearing and this porous bearing of a topped part.This reaction medium comprises a conversion zone at least, and preferably, comprises that is at least checked a zone, and wherein, this checks the zone can provide the pattern that is suitable for differentiating a checked object.In a preferred embodiment, this checks the zone can provide a kind of pattern, and this pattern comprises the copying surface of other part of the health of an impression of the hand or a checked object that can be used for differentiating.Above-mentioned conversion zone comprises a kind of device that is used to carry out ligand/anti-ligand or ligand/ligand-receptor determination, preferably, is a kind of device that has or not the mensuration of certain analyte.In another preferred embodiment, the device that is used to measure comprises the member that a kind of immunity is right.

An alternative embodiment of the invention is at a kind of determinator that comprises a kind of reaction medium, this reaction medium has a conversion zone at least, and preferably, has a zone of checking that comprises a member that a kind of immunity is right at least, wherein, this checks the identity that the zone can confirm a checked object.Preferably, the described zone of checking can provide a kind of pattern, and this pattern comprises an impression of the hand or the copying surface of the other part of the health of the above-mentioned checked object that can be used for differentiating.

According to a further aspect, the present invention includes preferably porous bearing of a bearing; Also comprise a kind of reaction medium that is installed on this bearing, preferably a kind of porous medium, wherein, described reaction medium has a conversion zone at least, and preferably, has at least one to check the zone, and wherein, at least a portion is checked the identity that the zone can confirm checked object.A preferred embodiment comprises and is arranged in an above-mentioned reaction medium or a right member of a kind of immunity on it.In another preferred embodiment, at least a portion is checked the member that the zone comprises that a kind of immunity is right.In certain embodiments of the present invention, described reaction medium can be no bearing.

The present invention is also at a kind of method that is used to handle sample, and this method comprises that a conversion zone that makes a determinator contacts with a check sample; Determine in described check sample, to have or not at least a analyte; And check the check pattern that forms a discriminating object in the zone for one at described determinator.

The present invention is also at a kind of method that is used for determining to have or not at a sample certain analyte, this method is included in the immunoassays of carrying out analyte in the conversion zone of an immunoassay apparatus, thus, determine in a check sample, to have or not certain analyte; And check the identity that the zone confirmation provides the people of above-mentioned check sample at one that is arranged on the described immunoassay apparatus.

Method of the present invention is included in determines to have or not certain analyte in the sample, preferably use any mensuration based on the ligand combination, and confirmation provides the people's of this sample identity, preferably uses any record based on immunity.With reference to Fig. 1, can determine in a sample, to have or not certain analyte by a kind of biofluid being applied at least one conversion zone 20 and making the association reaction of predetermined ligand/acceptor to take place.In a preferred embodiment of the present invention, finish described reaction and can cause producing a signal.Preferably, this signal produces with visible color or the form of not having a color.For example, in Fig. 2, the generation of having been drawn the zone 32 to 34 of hatched visible color shows no described analyte in this check sample, and achromatic region 35 to 37 shows that described analyte is arranged in this check sample.The ad-hoc location of specific type, the positive and/or the negative findings in conjunction with the method for measuring, produce above-mentioned signal, the signal that produced of not planning that the present invention is limited in and being adopted.

According to the present invention, can (for example differentiate body part by applying one, a people's finger or pin) check the zone to one, and make a predetermined ligand/receptor response to take place, produce the image of an impression of the hand or footprint thus, finish producing image or the pattern that a discriminating provides the people of this check sample like this.In a preferred embodiment of the present invention, a people's finger tip is coated with a kind of reagent that produces signal, preferably a kind of reagent that forms color, this finger tip is applied to a zone of checking with MIP, and this MIP can combine with the reagent of generation signal on the finger that is coated in that people.In a preferred embodiment of the invention, finish the image that the immune response meeting causes producing a visible color, this image is corresponding to that people's impression of the hand.Fig. 2 shows typical, a visual impression of the hand.

In one embodiment of the invention, especially for paediatrics, image that can form or pattern can be the parts of pin or hand or pin.In this embodiment, being coated with a kind of pin of reagent of tape label or the part of pin in advance can be applied on the device of the present invention.

Referring now to description of drawings exemplary embodiments of the present invention.

An immunoassay apparatus 10 of the present invention comprises a kind of reaction medium 14 that is preferably mounted on a bearing or the substrate parts 12.Preferably a kind of porous bearing of substrate parts 12.In a preferred embodiment of the present invention, reaction medium 14 comprises a conversion zone 20 at least and comprises that is at least checked a zone 16.In those exemplary embodiments illustrated in figures 1 and 2, immunoassay apparatus 10 comprises that six conversion zones 20 (label 32 to 37) and two

check zone

16 and 18.

In a pick-up unit 10 of the present invention, at least one in those conversion zones 20 comprises the device that is used to carry out immunoassays, has or not certain analyte so that preferably confirm in a check sample.In the exemplary embodiments shown in Fig. 3, conversion zone 20 comprises a kind of carrier that is embedded in the reaction medium 14 at least, for example, and particle 22.As illustrated, with a kind of immunity a member of 21 is coated on particle 22.

In a pick-up unit 10 of the present invention, check the device that at least one in 16 of zone comprises the identity that is used to confirm a checked object, this checked object preferably includes a right member of a kind of immunity.In exemplary embodiments shown in Figure 4, check zone 16 and comprise a kind of carrier that is embedded in the reaction medium 14, for example particle 24.As illustrated, with a kind of immunity a member of 25 is coated on particle 24.In a preferred embodiment of the present invention, MIP25 can be attached on a kind of reagent that produces signal, or is attached on the reagent couplings that a kind of MIP-produces signal.

By reference Fig. 5 to Fig. 8 typical method of the present invention can be described.In Fig. 5, can determine to have joined in the check sample of a kind of MIP30 one has certain analyte 31.Biofluid is applied on the surface of the conversion zone 20 with particle 22, particle 22 is with a kind of immunity member of 21 to be applied.If analyte 31 is arranged in the described biofluid, it will be attached to MIP30 or be caught by MIP30, mix with it and cultivate it with MIP30, and MIP30 be specifically designed to analyte 31.The immunity that finally obtains can not be attached to MIP21 to 50 or be caught by MIP21, but can pass through reaction medium 14.According to this embodiment of the present invention and in this case, the reagent that does not form color can be attached on the coated particle 22.

In Fig. 6, can determine to be added in the check sample of a kind of MIP30 one does not have certain analyte.Biofluid is applied on the surface of the conversion zone 20 with particle 22, particle 22 is with a kind of immunity member of 21 to be applied.In above-mentioned biofluid, do not have analyte, MIP30 can by MIP21 combination or catch.According to this embodiment of the present invention and in this case, the lip-deep a kind of reagent that forms color that is applied to reaction medium can be attached on those MIP30 on the described coated particle.In an alternative embodiment of the present invention, MIP30 can comprise the reagent of above-mentioned formation color.

Fig. 7 and Fig. 8 have illustrated at one and have checked a kind of typical method that produces the image of an impression of the hand on the regional surface.A people's finger surface 60 comprises some crestal line 61 and some ditch lines 62.When on the finger that applies a people with a kind of reagent that forms color 102, the reagent of the described formation color of major part can be tending towards collecting in those ditch lines 62.When this finger is applied to reaction medium 14 surperficial, can believe that the zone of the reagent of the formation color of higher concentration is arranged in the zone of described those ditches.Coated particle in the surface of checking the zone generally can be attached to the reagent of the described formation color of higher proportion, and this reagent is transferred a kind of stronger visible color of generation corresponding to the ditch line of above-mentioned people's finger.

In a preferred embodiment of the invention, generally only in a few minutes, just finished and determine in a kind of biofluid, to have or not certain material and produce an impression of the hand image.

Quick and the higher susceptibility of apparatus and method of the present invention promptly is applied to the device and the mode on film surface to reagent and/or check sample, and to a certain extent, forms those materials of described film all owing to following two characteristic.Like this, as described below, the mode that applies and material make those reagent in the solution keep contact with described film surface one period of length, so as promotion greatly, the reaction between those reagent in described film surface and those reagent in solution.The reagent that this just causes higher susceptibility and needs low concentration.

To illustrate in greater detail various aspects of the present invention now.

Ligand-receptor determination

According to the present invention, can adopt the pick-up unit that uses with any determination techniques that is used to detect analyte of interest.In the mensuration of the present invention of all uses, a kind of analyte is at least in conjunction with a kind of ligand or ligand-acceptor.Typical in measuring the ligand assay that includes, but are not limited to those tape labels, these mensuration comprise those competitive binding assays of a kind of solid-state binding reagents that is coupled, for example adopt the cultivation simultaneously or sequentially of one or more independent steps, and also comprise determining displacement; And, typical in the mensuration of measuring the binding reagents that also includes, but are not limited to those tape labels, these mensuration comprise noncompetitive and emulative in conjunction with measuring, it is those mensuration and the sandwich assay of binding reagents or ligand mutually that this mensuration comprises wherein solid-state, and this sandwich assay comprises the immunosorbent assay of precipitation, radiommunoassay or ligase.At this specific record of not planning to make the present invention be limited to the type of used immunoassays or being used to carry out this mensuration.Typical immunoassay has been shown in some examples.

Reaction medium

According to the present invention, reaction medium can be any medium that is suitable for carrying out ligand-receptor determination.As mentioned above, reaction medium comprises a conversion zone at least, so that show and brightly have or not certain analyte, and preferably comprises one at least and checks the zone, so that show the identity of bright checked object.In general, ligand-receptor determination can adopt the reaction medium of following form to carry out, and this form is pearl, pipe, plate, wheel blade, a kind of porous or does not have pore matrix or porous or non-porous film or filtrate.

Reaction medium can comprise a kind of medium of thickness, and wherein, a zone of surface or this near surface comprises a conversion zone at least.Reaction medium also can comprise a kind of rhythmo structure that makes a conversion zone as a superficial layer.According to the present invention, react and check the zone and be arranged in reaction medium, thereby make and to be easy to visual a kind of reagent (if any) that produces signal.

According to the present invention, preferred reaction medium is a porous, and even more preferably, be a kind of perforated membrane.Do not plan to make the present invention to be limited to the type and the structure of described reaction medium at this.Those of ordinary skill in the art is easy to recognize that the selection of reaction medium depends on physical properties desired, this physical property comprise but and be limited to following these character, for example, susceptibility, the type of binding ability, stability, molecule or the type of combinative those MIP and the compatibility that writes down with certain special mensuration.

The typical material that is used to form reaction medium includes, but are not limited to latex, nitrocellulose, nylon, cellulose and nylon.

A kind of preferred reaction medium is commercially available Gelman Super board film.Super board film is the polysulfone membrane of a kind of possess hydrophilic property surface, low protein combination latex.A those of ordinary skill in the art will appreciate that may need this film, because it provides good flow velocity and particle hold-up; Smooth surface; Pure white and opaque, so that enhancing signal contrast in certificate test; Extract is few, so that reduce sample contamination and background interference; And uniform factor of porosity, so that guarantee the consistance of final products.Adopt this film also to eliminate needs, may need this wetting agent, so that undesired extract is introduced in control outside wetting agent.

Preferred film in addition, the IAM motion suppresses compatibility film (ImmobilonAffinity Membrane) and (can buy from Millipore company on the market, for example, see U.S. patent documents US 4,407, No. 943), the hydrophobicity substrate polymer, polyvinylidene fluoride (Polyvinylidene difluoride) (PVDF), conjugated protein.IAM also makes the scope can highly control whole protein bound, and the user can repeatedly fix the amount of protein from the teeth outwards from the nanogram to the microgram, so that the needs of suitable various mensuration.Road known in the art is incorporated into IAM and its purposes to a kind of ligand.

Mobile characteristic of passing in view of the additional facility of solid-state phase, typical high protein binding ability and film may adopt some films as the solid bearing in reaction medium of the present invention.Nitrocellulose is owing to its high compatibility for protein nucleic acid, some other cellular antigens and some cell macro molecules, it is one of film of the most common employing, and, if the combination of MIP is words ion-type or hydrophobic, may need nitrocellulose.Nitrocellulose also provides a kind of good matrix for erase protein and nucleic acid.Nitrocellulose can be cut into desired Any shape, and has useful like this characteristic, and promptly the amount of the protein in the impression of the hand is clearly visible.

On film, change or add different surface naturies and comprise within the scope of the invention in order to obtain needed result, for example, be designed for surface nature for the film of the competitive binding assay of hormone and can be different from immune measuration method for certain medicine.For example, already through showing, with a hydrophiling of ethanolamine treatment pvdf membrane reduced the non-specific bond on film surface.The surface treatment reagent of certain special pearl or the selection of surface nature can be based on giving this surperficial chemical characteristic; Make on conversion zone or the functionality sex change of bioactive agents wherein or the Disability or the reduction of giving this functionality; Realize the requirement of certain orientation of the bioactive molecule that certain is fixed; Promote the requirement of the long-time stability of the bioactive molecule that certain is fixed; Comprise certain needed nucleophilic substitute; And the availability of reagent treatment and cost.Adopt some other surface treatment reagent, comprising difunctional or poly functional reagent, so that influence the surface nature of film, these all comprise within the scope of the invention.

Those of ordinary skills are by comprising that into a kind of reaction medium of porous is easy to recognize the benefit of being brought.Positive check sample will be easy to by reaction medium, eliminate the needs of cleaning reaction dielectric surface before adding the reagent that forms color.Although lessly be ready like this, but plan to make to the present invention includes non-porous medium.A kind of preferred seat material is the porous plastic materials that a rigidity is arranged of a kind of Porex of being called.

As mentioned above, reaction medium can be by bearing or not by bearing.In a preferred embodiment, reaction medium is by bearing, and even more preferably, this bearing is a porous.A kind of typical bearing includes, but are not limited to polystyrene.

In another embodiment of the present invention, bearing can be can not permeate fluid and when testing used those reagent; In this embodiment, reaction medium preferably " thick " to medium the surface or its near be enough to have a conversion zone.

In preferred embodiments more of the present invention, wherein said device all or is substantially provided for oneself, promptly need not other reagent or only come the cleaning reaction medium at the most with several buffer solution or water, and the technology that only needs floor level is finished accurate mensuration, and the material of preferred another kind of type comes the bearing reaction medium.For more such purposes, the material of preferred a kind of compressible core or similar sponge.Like this, by being exerted pressure in vicinity, the flat basically surface of outside of reaction medium that contacts or be attached to a flat basically surface of compressible seat material, cause compression or load mould, thereby cause abutment surface to be lower than its not compressed flat surface on the surface of propping up seat material of contiguous reaction medium.When having certain liquid, after removing pressure, and expand into it not during compressed state when this compressible material from compressible seat material, described liquid is drawn into described seat material.Like this, if a kind of fluid sample, reagent solution, buffer solution or water are applied on the reaction medium, and apply enough pressure, to cause the distortion in the surface of a seat material, liquid is drawn in this seat material when described seat material expand into its virgin state.Institute's applied pressure is a compressible function of described material.A kind of preferred compressible seat material is a kind of compressible, soft, webbed isocyanurate foam, preferably, is a kind of polyester or polyether-polyurethane foam with pore rating of about 75 to 100ppi, and preferably, pore rating is 80 to 90ppi.Some materials like this can buy from E.M.Murray incorporated company.

Check the zone

In a preferred embodiment of the present invention, reaction medium comprise one be used for when determining that a check sample has or not certain analyte, establishing a reference point check zone (with reference to checking the zone), and the most important thing is a checked object that is used to confirm to provide described check sample identity check zone (identity is checked the zone).

According to the present invention, be used to confirm to provide of identity of the checked object of check sample to check zone (identity is checked the zone) and can comprise that any signal that combines a kind of MIP sends mechanism.For example, check the member that the zone can comprise that a kind of immunity is right, this immunity is to can be in conjunction with a kind of reagent that produces signal, when the finger by the exposure test object reagent of this generation signal is applied to identity check the zone the surface on the time, just on verifying attachment the generation a described finger image.

As used herein, " can confirm the identity of checked object " is meant and specifically identifies the ability that the object that is verified sample is provided.In a preferred embodiment of the invention, check the member that the zone comprises that a kind of immunity is right, when placing this immunity when the finger with the checked object that has scribbled a kind of reagent that produces signal is in advance contacted, this immunity is to producing a kind of permanent image of an impression of the hand, preferably, be a kind of visual impression of the hand image that is easy to.In another embodiment, the present invention includes one have a kind of immunity to or a member of nucleotide acid sequence check the zone, this nucleotide sequence is can be in conjunction with the nucleotide sequence that can be used to the diagnostic test object.

According to the present invention, the zone of checking that is used for establishment reference point when determining that check sample has or not certain analyte comprises any apparatus that is used to provide the standard of checking.Show the several examples that are used to provide the standard of checking in some instances.At this, do not plan the present invention to be interpreted as being confined to be used for establishing the device or the mechanism of standard of comparison when differentiating assay.

Conversion zone

Conversion zone is meant the position that can determine to have or not certain analyte.For example, conversion zone preferably on the surface of reaction medium or near it, perhaps, can be under this surface.In a preferred embodiment of the invention, conversion zone comprises a kind of ligand or is attached to a kind of particle in certain matrix or medium or the ligand-acceptor of bead that this ligand or ligand-acceptor are suitable for catching certain analyte of interest at least.In another embodiment of the present invention, at least one in above-mentioned those conversion zones can be a kind of thing of checking that is used to establish a reference point, for example, a kind ofly will show the colourless thing of relatively checking.

Produce the reagent of signal

The reagent that produces signal is meant certain detectable signal of any generation or makes the reagent or the label that can detect certain ligand or ligand-acceptor.The preferred reagent that produces signal is those those reagent that need not with instrument, preferably just can detect with visual means analyte.The typical reagent that produces signal includes, but are not limited to the reagent that those form color, for example, and enzyme, the polymkeric substance that contains dyestuff, chemiluminescent reagent, fluorescent reagent, radioactive isotope or ferromagnetic particle thing.The reagent that forms color can be that a kind of colored particle, a kind of colored molecule or some can excite a series of materials that cause forming the incident of certain colored label, for example enzyme.Colored molecule can be a kind of fluorescent dye, for example fluorescein or rhodamine; A kind of chemiluminescent compound; A kind of noctilcent compound; Or a kind of compound that detects by absorption of electromagnetic radiation (and may launch again with another wavelength), described electromagnetic radiation comprises UV radiation, visible radiation and infrared radiation.Colored molecule can be directly or indirectly and certain ligand or ligand-acceptor phase conjugate.Can be selectively, colored molecule can be combined into a kind of particle, particularly a kind of particulate.

Be used for comprising alkaline phosphatase, horseradish peroxidase or B-galactosidase as the reagent that forms color.Some enzymes so often use with the substrate that adds lustre to.In example 11, provided the table that can form some substrate that typically adds lustre to that reagent uses below with some enzyme colors.

In a preferred embodiment of the invention, the reagent that forms color can be the dense particle of a kind of electronics, for example, and collaurum, silver-plated collaurum or ferritin.

In another embodiment of the present invention, the reagent of formation color can combine with a kind of MIP.

The present invention includes a kind of reagent that produces signal, this reagent can be the reagent that produces signal.Preferably, the reagent that produces signal comprises and is used for detecting the absorption at the conjugate of reaction medium or certain analyte on it or analyte by a kind of MIP of tape label.The MIP of tape label can comprise any one of a large amount of suitable marks, for example, and enzyme, fluorescent chemicals, noctilcent compound, ferromagnetism atom or colored particle or particulate.In a preferred embodiment, being used for mark of the present invention is collaurum.Gold is biologically inert, good CHARGE DISTRIBUTION is arranged and can many useful forms be widely adopted.Can strengthen its detection with the method for several depositing silvers, these methods make can use bore hole monitoring development situation.The colloid gold particle thing of available AIA, a-protein or the streptavidin conjugation with broad range in market.

Although a kind of collaurum substrate is better than the particle or the particulate of other dyeing, and a kind of substrate that adds lustre to provides a kind of enzyme conjugate that is used for, alternative sensitive detecting method.

Several embodiments of the present invention relate to pocket determinator and analyze box, this device and analyze box comprise all or most for qualitative and/or quantitatively determine necessary those reagent of one or more analytes and material.A preferred embodiment of such device has been shown among Figure 11, and Figure 11 has shown a kind of determinator of providing for oneself 110.This device comprises a shell 111, and in a preferred embodiment, shell 111 comprises one first parts 111a and one second parts 111b.A kind of reaction medium 114 of the above-mentioned type is arranged in described first part of case shell 111a, is installed on the upper surface of step that an of substrate parts 112 raise or terrace part.Substrate parts 112 preferably includes a vicinity, contacts or be attached to the porous bearing of reaction medium 114.In a preferred embodiment of the present invention, reaction medium 114 comprises one or more conversion zones 120, each conversion zone 120 comprises a kind of specific reagent, for example, (it can be a right member of a kind of immunity to a right member of a kind of ligand/acceptor, for example, a kind of antigen or antibody).Reaction medium in this preferred embodiment comprises that is at least checked a zone 116.As among those embodiment that discussed in the above, each above-mentioned conversion zone comprises that a kind of being applicable to determine the qualitative existence that certain is analysed thing or the reagent of quantitative scope.In a preferred embodiment of the present invention, check the zone for one and be applicable to giving and the person of discriminating one check sample.Equally, check zone 116 and also comprise a kind of suitable testing reagent that is used for the diagnostic test object, the member that preferably a kind of ligand/acceptor of this checked object is right for example, is a right member of a kind of immunity.

Those reagent that exist in each conversion zone can manually apply, or preferably, adopt a kind of automated method, for example, employing is used for a kind of ink-jet sprinkler of the sort of type of ink-jet printing system, so that described reagent is applied in single people and those conversion zones of keeping apart.

A kind of medium 126 is arranged in second parts 116 of shell, is scattered with the reagent of at least a generation signal or the reagent of indication analyte on medium 126, preferably a kind of reagent that forms color of this reagent.The medium 126 that contains the reagent that produces signal by factor of porosity than reaction medium or be used to hold that the lower material of medium of a check sample (the following discussion) forms.The medium that is scattered with the reagent that produces signal above preferably being used as is a kind of polyolefine material of closed porosity, for example, and the tygon of closed porosity or polypropylene.A kind of preferable material can have been bought as microcell from Voltek incorporated company.As used among other those embodiment of the present invention, the member of the tape label that preferably a kind of ligand/acceptor of reagent of generation signal is right, for example, the member of the tape label that a kind of immunity is right, for example a kind of material of colloid gold label.The medium 126 that contains the reagent that produces signal is positioned among the second parts 111b of shell in such a way, promptly when two members closes of shell when being in a specific detent position together, make between medium 126 and the reaction medium 116 to produce contact, thereby the material transfer that makes the generation signal that is included in the medium 126 is in above-mentioned reaction medium.Preferably, finish this point on 128 by medium 126 being installed in the bottom, a similar table top or the platform that has raise in bottom 128, in preferred embodiment shown in Figure 11, bottom 128 has the shape of substrate of being similar to 112.In this setting, the medium that contains the reagent that produces signal one first or open position and reaction medium 114 separate, and one second or specific detent position the agent transfer of described generation signal to reaction medium 114, owing to when the described medium 126 that contains the reagent that produces signal contacts with reaction medium 114, applied pressure, the speed that this setting obtains higher susceptibility significantly and increased.Can adopt superatmospheric pressure, this depends on related specific material, preferably a shade below about 2psi to about 5psi, after medium 114 and 126 was placed or contacted with each other, experience one section was enough to the reagent of generation signal suitably is retained near the surperficial of above-mentioned reaction medium or its to about 20 seconds time in about 5 seconds.Although generally can adopt higher pressure, so that the conjugate (such as discussed below) of this analyte that shifts the reagent, analyte of above-mentioned generation signal or form with a kind of ligand acceptor of this analyte, the variation because pressure and time are inversely proportional to, the duration of exerting pressure reduces.

One other component that provide for oneself or " complete " device shown in Figure 11 is a kind ofly to be used to hold and to shift the medium 185 of a part of certain check sample to reaction medium 114.Be used for keeping a kind of check sample and can be a kind of hydrophilic material with the medium 185 of a kind of reagent of certain analyte reaction, preferably, be certain water wettability sponge form and the material that forms by a kind of micropore polyester urethanes sponge material.A kind of preferable material is a polyester polyurethane foam sponge material, and this material porosity is 100psi, and can buy from E.N.Murray company limited.Medium 185 is used for several purposes.At first, it can absorb and take from an a kind of check sample of giving with person or checked object.It can also transfer to reaction medium 114 to this sample or a kind of acceptor that is present in the formed a kind of conjugate of certain analyte in this sample and resists this analyte.Medium 185 can also for be present in those materials in the described check sample and be present in the above-mentioned medium a kind of reagent the two all provide the diffusion and reacting environment.The member that preferably a kind of ligand/acceptor of this reagent is right, for example, discussed above and in Fig. 5 and Fig. 6, differentiate a right member of a kind of immunity for the sort of type of MIP30.Reagent in medium 185 generally is a kind of antibody.This reagent is contained in the medium 185, preferably as a kind of freezing dried antibody.In this embodiment, the solution-treated of a kind of reagent of sponge medium 185 usefulness, and processed sponge is subjected to the freeze drying processing in a freeze drying chamber, and for example, in a Speed-Vac..

Preferably to be used to hold the end that the medium of check sample and this check sample and reagent can be installed in a right cylinder 140 by lyophilized form.In the embodiment shown in fig. 11, all the base part 128 than the case member 111b that the medium 126 that contains the reagent that produces signal is installed thereon is big with respect to the shape of the open end 140a of the above-mentioned cylindrical blind end 140b that medium 185 is installed thereon or diameter.Preferably, the shape of the open end 140a of right cylinder 140 is corresponding with the shape of base part 128, thereby this right cylinder can be easy to be contained among the case member 111b above base part 128.The shape of right cylinder 140 and structure make when above-mentioned those case members among medium 114 and 126 is in the relation that separates each other are in one " opening " position, and medium 185 is also opened or primary importance and medium 114 and 126 boths are in the relation that separates each other case member 111a and 111b above-mentioned.One second or " sealing " position at case member 111a and 111b, and remain in the shell 111 with the relation that bottom 128 and 126 one-tenth in the medium that contains the reagent that produces signal separate each other simultaneously, certain analyte that those materials in being used in the medium 185 that holds sample and being in this sample and/or be formed on are found in this sample and a kind of ligand/acceptor between its a kind of conjugate are to contacting and transfer to reaction medium 114.Like this, above-mentioned second or detent position, by the cylindrical end 140b and 185 application of forces of the medium on reaction medium 114, this force mechanism and be presented on the medium 126 that contains the reagent that produces signal and the way of contact that is between the above-mentioned reaction medium of one second detent position roughly the same.The method of using this device is discussed below.

Although Figure 11 has illustrated the one embodiment of the present of invention that adopt one two parts shell 111, this shell 111 has one first parts 111a and one second parts 111b, described embodiment also has dismountable parts 140, a kind of sponge 185 that is used to hold a check sample is attached to parts 140, but the present invention still considers others and some alternative embodiment.For example, case member 111a shown in Figure 11 and 111b are that hinge means with a softness is connected to each other together.When described shell is made of a kind of suitable plastic, these plastics are inertia to any reagent or the material in the solution of those media other element or that put on this device of being used for device, at this moment, above-mentioned hinge can be made by the same plastic material of a thinner part.Like this, above-mentioned shell can be made for an independent unit in a stamping procedure.Can be selectively, other material can be used as above-mentioned hinge material, and described hinge is connected to above-mentioned those case members with proper device.

In another embodiment, case member 111a need not to be connected to each other to be in the same place with 111b fully.Like this, stopping device, can become in place with a ratchet or some similar device interlock to those independent unit architectures, perhaps, those independent parts can engage one another with a threaded device or a bayonet joint.

In another alternative embodiment of the present invention, the medium 185 that is used for holding sample and holding a kind of testing reagent can be positioned at one the 3rd case member, rather than is contained on dismountable columnar component 140 of type shown in Figure 11.When those case members that permanently link together mutually use together, for example explanation just like that, contain the case member that is useful on the sponge that holds a kind of check sample and can be placed on those of the medium that holds reaction medium and contain the reagent that produces signal independently case member (being equivalent to case member 111a and 111b) is middle.But, in this example, the case member that contains the centre of sponge 185 can engage one or two in two other case member, and when in place and when forming the parts of this shell, contacts with reaction medium in first case member.When wanting reaction medium 114 is contacted with the medium 126 that contains the reagent that produces signal, can also remove it.

In yet another embodiment of the present invention, a hinge that contains on the marginal portion that the case member that is useful on the water wettability sponge that holds a check sample can be used on first case member is connected to this first case member (being equivalent to the case member 111a shown in Figure 11), and this marginal portion is relative with the edge of this first case member that is connected in second case member.Like this, when wanting to place the sponge that contains a check sample to such an extent that contact with reaction medium, the 3rd case member that contains this sponge can be rotated to a second place of the transferable material that contacts with described reaction medium, and after this can be rotated to one with above-mentioned reaction medium spaced positions from each other, and second parts of shell that contain the medium 126 of the reagent that produces signal may rotate to above-mentioned reaction medium and contact transferablely.

The above embodiment of the present invention that contains three kinds of media (a kind of reaction medium, a kind of medium that is used to hold a check sample, and a kind of be used to hold a medium that produces the reagent of signal) is provided for oneself basically.Like this, just need not additional other reagent in order to finish mensuration.At the most, use a kind of buffer solution in final step, for example a kind of phosphate buffered solution or water are so that the cleaning reaction medium, from those reagent of the surface removal remnants of this reaction medium.Can selectively when a kind of compressible porous medium is used as the bearing of reaction medium, can need not to use any additional materials.

The present invention also considers the pocket determinator of only using two media, and is not the device of above-mentioned three kinds of media.Like this, in the embodiment shown in fig. 12, although also keep reaction medium 114 to be installed in the substrate 112 in the first of shell, contain and the sponge 185 that shifts sample can be directly installed in the substrate 128, rather than the medium 126 that contains the reagent that produces signal is positioned at the there.In a such embodiment, can exempt right cylinder 140, in the embodiment of " three kinds of media " shown in Figure 11, medium 185 is installed on the right cylinder 140.In a such embodiment, can use the washer bottle that contains a kind of buffering agent, washing agent or water in the same manner as described above.But, in this embodiment, the reagent that produces signal can directly be distributed in from a reagent bottle on the finger or other check body part of checked object, thereby and be placed on the sponge 185 that is loaded in the substrate 128 at check sample, begun to cultivate and shell sealed between sponge 185 and reaction medium 114 with suitable pressure the contact of transferable material is arranged after, above-mentioned finger or other body part can be placed to such an extent that contact with reaction medium.

The device of another kind of two media has been shown among Figure 13.Resemble shown in Figure 11 and embodiment discussed above, this comprises reaction medium 114 and the medium that contains the reagent that produces signal 126 in second parts of shell in the substrate 112 that is installed in one first parts that are in shell.Except not having right cylinder 140 and the medium 185 that is used to hold a check sample, this corresponds essentially to the embodiment shown in Figure 11.The device of three kinds of media shown in the image pattern 11, this embodiment do not need checked object and exist, and therefore when checked object does not exist, can not produce impression of the hand or other discriminating mark of checked object.But, can selectively when testing, there be checked object on the scene, can produces impression of the hand.In this embodiment, a kind of reagent mix that sample and device are equipped with, this reagent is suitable for being loaded in the reagent bottle most, as the parts of analyzing box.Described reagent can comprise one or more materials, this material can show the existence of bright one or more analytes, and a right member of preferably a kind of ligand/acceptor of mentioned reagent, most preferably be a kind of immune right member that can form conjugate with a specific analyte.The potpourri of above-mentioned check sample and reagent drop by drop is applied to the surface of reaction medium, time will be enough to make cultivation to carry out, and the time of one section weak point of shell seal, be enough to of the lip-deep contact of the agent transfer of described generation signal to above-mentioned reaction medium thereby between medium 126 that contains the reagent that produces signal and described reaction medium, produce.If when check, have checked object to exist, not stopping device so that between medium 114 and 126 contact is arranged, but a finger of checked object is rolled on the surface of the medium 126 of the reagent that produces signal, then this finger is pressed on the reaction medium.

An alternative embodiment of the invention is the device and method that is used to distinguish acute and chronic drug abuser at a kind of.Like this, in a people's who uses medicine body, not only find to have the molecule of this medicine or certain metabolin, and this medicine or such metabolin are attached on a kind of carrier molecule.Constantly use certain medicine irritation to produce a kind of antibody of resisting the carrier of bound drug.Like this, in the people's who uses certain medicine for a long time body, not only can find the residue of this medicine or its metabolin, and can find to have human antibodies for described medicine.In an embodiment of apparatus of the present invention, can put a conversion zone in reaction medium to the some drugs of checked object use under a cloud point.Then, if in a check sample, exist, will form a kind of conjugate that has the human antibodies of said medicine as certain analyte.Can adopt corresponding goat or mouse to resist the antibody of human tape label as a kind of reagent that produces signal that is used to show that bright described drug antibody exists.

The method of using

A kind of assay method of the present invention comprises for certain analyte in a conversion zone of a determinator and carries out ligand-receptor determination, determine in check sample, to have or not certain analyte thus, and check the people's who confirms to provide described check sample in the zone identity of said determination device.In a preferred embodiment of the present invention, can adopt some ligand-receptor assays, for example, in a kind of reagent that produces signal, finish the people's who confirms to provide check sample identity in conjunction with a kind of MIP.

As mentioned above, on the surface that will be used to check the biologicfluid sample that has or not at least a analyte to be applied to immunoassay apparatus of the present invention.Can adopt described fluid sample is applied on any device of testing instruments.For example, in one embodiment of the invention, biofluid (for example, urine) can combine with a kind of MIP by the immunoassay of standard, is drawn into a suction pipe, and is deposited on the surface of verifying attachment.

In another example, biofluid (for example, blood) can be collected in the suction pipe and be deposited on the surface of verifying attachment, and perhaps can spread upon a solid and grant on the thing, for example, finger tip.In a preferred embodiment, blood is coated on the finger tip of checked object, then, this finger tip is pressed contiguously by the surface at pick-up unit.In another preferred embodiment, biofluid combined with a kind of MIP before being applied on the reaction medium.Also have among the embodiment, sample deposition is on the surface of the medium that is used to hold a check sample.In some instances, the other typical method that check sample is applied to pick-up unit has been described.

A preferred embodiment of the present invention comprises that working pressure is applied to one or more reagent or a kind of sample on the surface of verifying attachment.For example, found a kind of reagent that produces signal is spread upon on the finger tip of checked object, and this finger tip is applied on the verifying attachment, reduced the amount of the reagent of needed generation signal significantly, and increased susceptibility significantly and measured the speed of carrying out.For example, in order to finish mensuration, can be few collaurum to 10 microlitres, the collaurum of preferably about 12 microlitres spreads upon on the finger tip of checked object.By comparison, for the common mobile box determinator that passes, generally need as many as 100 microlitres or more produce the reagent of signal.In a kind of similar mode, another preferred embodiment is included in and has used pressure to sample when being applied on the surface of reaction medium.

Although do not plan to be confined to a kind of specific theory of operation, but pressure that is occurred when should adjacent finger being placed on the determinator and/or surface tension when checked object cause making ligand and ligand acceptor increase concentration localize, increased the time that those reagent remain on a regional area of determinator, and this may relate to the migration from the crestal line of impression of the hand to the ditch line of the reagent that produces signal, to produce a clearly impression of the hand pattern on the surface of verifying attachment.

Reaction more completely seemingly owing to taken place on the surface of the film that can find the reagent of having fixed in more hypersensitivity provided by the present invention and the amount of reagent that reduced.Reaction more completely obviously is owing to lip-deep those reagent that are applied to film with liquid form have kept longer a period of time there.Like this, as those the most common verifying attachments is typical, fluid sample and/or reagent are applied to dropwise form or contain reagent or contain on the surface of film of check sample, and be absorbed by this film soon, this is because a kind of absorbing agent contacts with described film on the downstream surface that is placed on it, or because of the character of described membrane material.But, in the present invention, the liquid that contains reagent and/or sample is applied on the surface of reaction medium, and by the pressure that applies greater than atmospheric pressure described liquid is remained there as a film, this film does not almost show the trend what will pass completely through above-mentioned film.Like this, adopt finger as applicator, or adopt other applicator, for example, as the included a kind of applicator of a part of device, the upstream face place that under pressure, shifts and remain on reaction medium at the liquid that contains reagent on the described applicator.Like this, although be reluctant to adhere to any theory, obviously, the initial surface tension of aforesaid liquid, and in conjunction with some other factor, for example, the capillary action of applicator, and may also have the factor of porosity of having placed the medium of aforesaid liquid in the above in conjunction with pressure, as long as on the outside surface of described medium, keep above-mentioned pressure, will cause the film of liquid to remain on the upstream face rather than pass described medium.Capillary contribution when a film is remained on reaction medium surperficial shows as and has increased those (those) that are present in the aforesaid liquid and plant the utilizability of material for some reagent in the described medium.Like this, adopt a kind of applicator, for example with finger, use is greater than the pressure of atmospheric pressure, generally be low to moderate the pressure of about 1psi, and be willing to a normal pressure that also cosily applies (for example, such by pointing by one " formal impression of the hand " time) so greatly up to a checked object to about 5psi, when removing described pressure, these have prevented that all aforesaid liquid from passing film.Pressure normally applies one about period of 5 to 20 seconds.Reagent is remained on the higher extent of reaction that such a period of time on the surface of medium or film has been enough to guarantee to be positioned at the upstream face place of described medium or directly has been in those reagent below it.Although the character of described liquid, for example its surface tension, and the material that constitutes above-mentioned film and applicator, with this phenomenon some relation is arranged, as seen, can adopt various materials, and the phenomenon that the aforesaid liquid film remains on the surface of described film still occur.Further, but also be to be unwilling to adhere to a certain specific theory, as seen, the raising of the determination test of apparatus and method of the present invention sensitivity and particularly rapidity to small part be owing to the time adopted pressure some liquid solutions being applied to the reaction medium surface.It is believed that institute's applied pressure is greater than atmospheric pressure and when generally being low to moderate 1psi, preferably when about 2 to 15psi scope, this pressure has just increased momentum or the speed that specific reaction takes place.

Verifying attachment preferably include one or more fixing in the conversion zone respective amount MIP.For example, if analyte of interest is a kind of antibody, above-mentioned verifying attachment can comprise a kind of microballoon imbedded in the conversion zone or anti-source of particle of being attached to.A those of ordinary skill in the art will appreciate that and can be fixed on the anti-source of requirement in the conversion zone, and the MIP of institute's combination of an available threshold concentration detects the analyte of a scheduled volume.In one embodiment of the invention, pick-up unit comprises several conversion zones, and each conversion zone is used for a kind of different analyte, and, each conversion zone comprises the MIP of institute's combination of a predetermined threshold value concentration, to detect the analyte of a scheduled volume.Such as used herein, threshold quantity or concentration are meant the lower limit for the concentration range of certain analyte.In another embodiment of the present invention, pick-up unit comprises several conversion zones, and each conversion zone comprises a kind of different threshold concentration to same analyte.In this embodiment, threshold concentration provides a reference point for the upper limit of the concentration range of definite certain analyte.As shown in those following examples, the threshold concentration that changes in above-mentioned those conversion zones can provide a kind of typical method of determining the amount of the analyte in the sample.

Be applied on the surface of verifying attachment at the biofluid that is verified after, can determine in check sample, to have or not analyte, preferably, determine a kind of color (signal) or do not have certain color (signal) by visual.In one embodiment of the invention, positive reaction is promptly shown the bright above-mentioned analyte that has, corresponding to colourless in described biofluid.In a preferred embodiment of the present invention, negative reaction, promptly show bright in above-mentioned biofluid no described analyte, corresponding to a kind of visible color.But, in one embodiment of the invention, determine wherein whether checked object is a chronic drug abuser, check a kind of human antibodies to certain medicine, and, the reagent that is present in the conversion zone is this medicine itself, and the reagent that has a generation signal of mark for example is a kind of anti-human antibodies with golden labelled goat or mouse, and brightly there is the positive reaction of said medicine antibody a kind of showing corresponding to color occurring.

According to one aspect of the present invention, when the biofluid that is verified is blood, be surprised to find that now whole blood can be applied directly on the surface of verifying attachment and need not separation of serum before sample is applied to this verifying attachment.According to this embodiment of the invention, whole blood sample is applied on the surface of verifying attachment after, a kind of washing agent is applied on the blood sample.Although people know, some washing agent, for example tween can be used to remove red blood cell and analog thereof from whole blood sample, be surprised to find that, apply removing reagent, for example, applying of a kind of washing agent (the preferably a kind of potpourri of ion detergent and nonionic detergent and washing agent of alcohol of comprising) make and can be applied to whole blood on the surface of determinator, and need not before being applied to described device isolated cell component from serum.Preferably apply a kind of washing agent, because this washing agent destroys haemocyte.Afterwards, ruinate cell debris passes above-mentioned medium and removes from conversion zone, promptly no longer disturbs the surface antigen fixed.

In case determined to have or not analyte, can differentiate the checked object that is verified biofluid with the pick-up unit of making according to the present invention.In a preferred embodiment of the present invention, the identity of determining checked object comprises carries out ligand-receptor determination, and this mensuration causes showing single people's impression of the hand.According to a preferred embodiment of the present invention, determine that the reaction of the analyte in checked object identity and any and above-mentioned fluid is irrelevant.

The method according to this invention, people's finger tip apply the reagent that a kind of and certain MIP carry out the generation signal of conjugation.When coated finger tip was applied on the surface of checking the zone, the conjugate of described generation signal was attached to a kind of MIP in the zone of checking that imbeds verifying attachment.Although the present invention is not limited to certain specific theory of operation, people should believe, those crestal lines of a people's finger tip and ditch line provide the regional area (corresponding to crestal line) of reagent of the generation signal of the regional area (corresponding to the ditch line) of conjugate of the generation signal that has concentrated and low concentration.When checking on the regional surface for one that is applied to a device of the present invention, just produce the image of the discriminating known of an impression of the hand, thus, can provide the people's that described check sample is provided identity.

According to the present invention, can be by determining in this sample, to have or not certain analyte of interest on the surface that sample is applied to a kind of reaction medium, described reaction medium comprises a conversion zone at least, and this conversion zone has a right member of a kind of immunity therein; And above-mentioned determine also will be by apply a kind of reagent that directly or indirectly is attached to MIP or is attached to the generation signal of above-mentioned analyte of interest (for example, a kind of secondary antibodies).

The verifying attachment of providing for oneself that can be as shown in the following Figure 11 of utilization:

A kind of check sample of liquid form, for example, a kind of body fluid (for example, saliva, urine or blood) is placed on the outside surface that is used to hold with the medium 185 of transfer check sample.Up to being applied to about 20 check samples on the medium 185.Because this device provides the high susceptibility and the speed of so Duoing that gets than those common verifying attachments, does not almost have a check sample preferably to be adopted, and but, preferably uses about 3 to 5 check samples.Make right cylinder 140 place substrate 128 top and in place at above-mentioned shell with its open end, and make the medium 126 that produces signal in place in the second parts 111b of shell, check sample is applied on the outside surface of medium 185, perhaps this sample is applied on described medium and the right cylinder 140, this is placed among the housing parts 111b.Any analyte that is present in the above-mentioned check sample has all experienced and the right member's reaction of a kind of ligand/acceptor that is present in lyophilized form in the sponge 185, and the member that preferably a kind of immunity is right is so that form a kind of conjugate.Allow to arrive about 4 minutes a period of time, preferably, about 2 minutes a period of time, be used for the right member's of above-mentioned check sample and ligand/acceptor cultivation (reaction) through about 1 minute.It is in place that right cylinder 140 is remained among the second parts 111b of above-mentioned shell, two parts of shell 111 are merged together to one first detent position, in this position, the outside surface contact reaction medium 114 of sponge 185 also applies enough pressure, so as to shift the right member of the above-mentioned ligand/acceptor of those materials be present in the above-mentioned check sample and be formed on analyte and member that described ligand/acceptor is right between any conjugate.In this position, although be called one " seal position ", described shell is not wholely to seal, and this depends on the specific structure that is adopted.Keep this detent position to arrive about 4 minutes a period of time, preferably about 2 minutes in about 1 minute.After this, open above-mentioned shell, and from canning, take out the right cylinder that accommodates sponge 185.Then, close a described complete closed or second detent position of installing, in this position, make the medium 126 contact reaction media 116 that contain the reagent that produces signal with enough pressure, so that shift the reagent of this generation signal, for example, a member of the band gold mark that a kind of ligand/acceptor is right is to described reaction medium.Make above-mentioned shell remain on this second or the position of complete closed at least about 5 seconds a period of times, preferably about 15 seconds a period of time to about 30 seconds.

After about one minute, by showing a white space in the suitable conversion zone, device will show the bright a certain amount of analyte that exists any analyte that is verified or existence to surpass a predetermined concentration or scheduled volume.In the overwhelming majority uses, when the amount that does not exist certain specific analyte or this analyte to exist makes that concentration is lower than a certain predetermined value, by the negative check of colour developing effect reflection of appropriate area.Device can not provide indication about the identity of checked object in this stage, unless other that this checked object is placed a finger or health after opening shell for the second time differentiates that part contacts with the zone 116 of checking of reaction medium 114.Can be selectively, making between medium 126 and 114 zone of checking that above-mentioned finger or other body part is not applied to described reaction medium after producing contact, but save coarctate this step of above-mentioned two media, and containing for the first time at the finger of above-mentioned checked object or other body part roll across on the medium 126 of the reagent that produces signal after, perhaps after the reagent that directly applies described generation signal with an applicator or distributor arrived above-mentioned finger or other body part, the finger of described checked object or other body part can contact above-mentioned reaction medium under pressure.Although differentiating in those methods of this checked object each by a kind of nonvolatil and individual characteristic of checked object all is fast and (the whole mensuration generally otherwise above about 3 to 5 minutes) of inspiration, in the end in Shuo Ming the method, the reagent that produces signal is applied directly on the finger tip, this finger tip is applied on the reaction medium then, the method of this last explanation is perhaps more effective, this to need to be the reagent and the sample of less amount, and the speed of this process perhaps more hurry up.Do not place method that a finger or other differentiate that body part contacts with reaction medium owing to thereby the speed efficient of required quantity of material and process is minimum, although between above-mentioned finger or other body part and transfer medium to contact effect medium.

The check analysis box

In one embodiment of the invention, the device of making according to the present invention can be assembled into an analysis box.This analyzes box also can comprise any of those reagent of being used for carrying out ligand/receptor determination in a large number.For example, described analysis box can comprise a verifying attachment, at least a reagent that is used to produce a kind of signal (or a kind of be used to shift the reagent of this generation signal " ink " pad to finger) and one or more flushings, cleaning or washing reagent.Described analysis box also can comprise one or more ligands or ligand acceptor, for example, freeze-drying primary antibody and a kind of secondary antibodies, and can comprise one or more buffering agents.Described ligand or ligand acceptor can be a kind of materials of tape label.

When above-mentioned those pocket devices were provided with the form of analyzing box, they can have the rinse solution that contains phosphate buffered solution.Embodiment to the two media that substitutes of the Figure 12 of above-mentioned discussion and device of the present invention shown in Figure 13 can provide such washing bottle and/or reagent bottle most effectively with a kind of form of analyzing box.Like this, the device that contains a shell and three kinds of media 114,126 and 185 that need not be shown in Figure 11, but adopt the device that comprises the two media in a shell and the above-mentioned three kinds of media.

The example that some are concrete

Example 1

An embodiment who prepares immunoassay apparatus of the present invention according to following method.By imbed a part of low-protein of being installed on the porous bearing film (Gelman Super film) with the granules of polystyrene thing of human serum albumin-benzoylecgonine (HSA-BE) coating, construct a conversion zone in conjunction with polysulfones.Check zone 16 by constructing on the different piece the polyethylene particle thing that applies with the anti-ageing murine antibody of goat being imbedded above-mentioned film.Above-mentioned reaction has been shown among Fig. 3 and Fig. 4 and has checked the typical schematic cross section in zone.

Prepare the said determination device according to following method.At first use a kind of retardance buffer solution (solution A) cleaning reaction medium, this solution is made up of 2% polyvinyl alcohol (PVA) (PVA) in the brine solution of phosphate-buffered, 1% glycocoll and 0.05% Tween-20.Mark the center well (check zone 16) of immunoassay apparatus with a kind of dilute solution of polystyrene latex, the anti-ageing rat immune globulin G of goat (absorbed anti-human serum) that this polystyrene latex is used in the salt solution of phosphate-buffered of the azide that contains 4% sucrose, 1.0% BSA and 0.05% applies.Dilute solution with a kind of polystyrene latex marks described conversion zone, and human serum albumin-benzoylecgonine (HSA-BE) that this polystyrene latex is used in the 0.2M sodium bicarbonate applies.Then described reaction medium is upside down on the hydrophobicity tygon, and was set in the hothouse between 80 and 100 dry one hour in temperature.After removing described reaction medium from above-mentioned hydrophobicity tygon, the said determination device just is ready to for using.

Example 2

To be verified from one and to have or not the person of benzoic acid ecgonine to obtain saliva sample.Aliquot 200 microlitres of the dilute solution of the anti-benzoic acid ecgonine of the mouse in the salt solution of phosphate-buffered immunoglobulin G (mouse anti--BE IgG) are added in the aliquot of above-mentioned saliva sample of 200 microlitres, described dilute solution contains 0.1% bovine serum albumin(BSA) (BSA) and 0.05% Tween-20.Allowing finally to obtain potpourri cultivated 3 minutes.Between this section culture period, the solution A of 400 microlitres is added in the reaction medium of immunoassay apparatus prepared in the example 1, and makes the solution A of this 400 microlitre discharge by this reaction medium.Then, mouse anti--BE IgG/ saliva mixture is added on the above-mentioned reaction medium, and cultivation can be carried out two minutes.Then, the aliquot of the solution A of other 400 microlitres is added on the above-mentioned reaction medium, and the aliquot of the solution A of these 400 other microlitres can be discharged.After solution A discharging termination, the goat that is used in the collaurum conjugation in the TRIS buffering agent of the azide that contains 1.0%BSA, 0.05% Tween-20 and 0.05% is anti--and dilute solution (being called " golden label solution " later on) 15 microlitres of mouse immunoglobulin G are coated on that people's that above-mentioned saliva sample is provided the finger.The reaction medium that makes this finger withstand above-mentioned immunoassay apparatus is slowly depressed, and is held in place for 3 seconds.Then, this is pointed careful ground roll from described reaction medium.Before being added to the aliquot of the solution A of other 400 microlitres on this reaction medium, make this reaction medium can cultivate for 15 seconds.After finishing this program, conversion zone colour developing shows negative assay for benzoylecgonine (for example, shown in conversion zone among Fig. 2 32 to 34 like that).

Except just add mouse anti--add before the BE IgG solution more a spot of benzoylecgonine to saliva sample, repeat said determination.In this example, after finishing check, above-mentioned those conversion zones are white (for example, shown in the conversion zone among Fig. 2 35 to 37 like that) fully.Finish two when measuring, center well is checked a marking (for example, shown in the impression of the hand 62 on checking zone 16 among Fig. 2 like that) of that finger that the zone comprises that people that above-mentioned saliva sample is provided.

Example 3

Method according to example 1 prepares immunoassay apparatus, except this device comprises 6 conversion zones that are used for different analytes.Each conversion zone comprises some granules of polystyrene things, and these granules of polystyrene things apply with a kind of different analyte conjugate that is embedded in the film.Conversion zone 32 to 37 comprises some polystyrene latexs, and these polystyrene latexs apply cocaine (32), some opiates (33), PCP (34), amphetamine/Corvitin (35), tetrahydrocannabinol (36) and ethanol (37) respectively as analyte (as shown in Figure 1).As example 1, with some be coated with goat anti--mouse immunoglobulin G, the granules of polystyrene thing imbedded in the film prepare central nucleus to zone 16.

Have or not a people of six kinds of analytes to obtain saliva sample on one's body from being verified.Each the mouse in the salt solution of phosphate-buffered for six kinds of different analytes anti--aliquot 200 microlitres of the dilute solution of analyte immunoglobulin G (being called " mouse anti--analyte IgG solution I " later on) join in the above-mentioned saliva sample of 200 microlitres.Described in example 2, when cultivating this potpourri, with solution A pretreatment reaction medium.Above-mentioned mouse anti--after analyte IgG solution I/saliva mixture is added on the described reaction medium, carry out the check program of the remainder described in the example 2.Handle above-mentioned reaction medium with solution A, with the goat that has scribbled the collaurum conjugation anti--a described reaction medium of finger contact of the checked object of the dilute solution of mouse immunoglobulin G, and and then handle this reaction medium with solution A.After having carried out this program, central nucleus contain the marking of that finger of that people that above-mentioned saliva sample is provided to zone 16.Conversion zone 32 all develops the color to 37, shows for a cloudy assay that has all six kinds of analytes.

Then, suppress remaining saliva with more a spot of amphetamine/Corvitin, tetrahydrocannabinol and ethanol.Then, mouse anti--aliquot 200 microlitres of analyte IgG solution I join in 200 milliliters the saliva of inhibition, and, repeat said determination as described like that for the sample that did not suppress.After having carried out described program, determinator has been showed the result shown in Fig. 2.Central nucleus comprise the marking of that finger of that people that above-mentioned saliva sample is provided to zone 16.Conversion zone 32 to 34 has shown negative assay, and all develops the color.Conversion zone 35 to 37 has shown positive result, and all is white.

Example 4

Method according to example 1 prepares a kind of immunoassay apparatus that comprises 6 conversion zones.As in the example 3, conversion zone 32 to 37 comprises some polystyrene latexs respectively, and these polystyrene latexs apply as analyte with cocaine (32), some opiates (33), PCP (34), amphetamine/Corvitin (35), tetrahydrocannabinol (36) and ethanol (37).But, in this example, the central nucleus prepared as Fig. 1 comprise some granules of polystyrene things to zone 16, and these granules of polystyrene things resist-coating of goat immunoglobulin G with the mouse that is embedded in the film.Have or not a people of six kinds of analytes to obtain urine samples on one's body from being verified.The mouse of aliquot 200 microlitres of this urine and 200 microlitres anti--analyte IgG solution I mixes the program of strictly testing then described in example 2.After measuring, the marking that central nucleus comprise that finger of that people that above-mentioned urine samples is provided to zone 16, and also conversion zone 32 all develop the color to 37, shows the negative assay for all 6 kinds of analytes of existence.

Then, suppress remaining urine samples with more a spot of amphetamine/Corvitin, tetrahydrocannabinol and ethanol.Afterwards, mouse anti--aliquot 200 microlitres of analyte IgG solution I join in the urine of inhibition of 200 microlitres, and repeat said determination as described like that for the sample that did not suppress.After finishing mensuration, immunoassay apparatus has been showed the result shown in Fig. 2.Central nucleus comprise the marking of that finger of that people that above-mentioned urine samples is provided to zone 16.Conversion zone 32 to 34 colour developings (negative findings), and conversion zone 35 to 37 is white (positive findingses).

Example 5

The reaction medium 83 for preparing the immunoassay apparatus shown in Fig. 9 and Figure 10 according to the method described in the real row 1.Reaction medium 83 has 6 conversion zones 32 to 37, these 6 conversion zones comprise some polystyrene latexs, these polystyrene latexs are coated with cocaine (32), some opiates (33), PCP (34), Corvitin (35), tetrahydrocannabinol (36) and ethanol (37) respectively as analyte, and reaction medium 83 also has central nucleus to zone 16, central nucleus 16 contain some granules of polystyrene things to the zone, these granules of polystyrene things are coated with mouse anti--goat immunoglobulin G, are embedded to (as shown in Figure 1) in the film.

A sterile swab 85 is placed on below a people's the tongue, checks this person's saliva to have or not six kinds of analytes.After one minute, take out swab 85 and be placed on the culture tank 80 of immunoassay apparatus and ware 81 in (as shown in Figure 9).The dilution of three silver is added on this saliva swab, described silver be coated with for the mouse of each of above-mentioned six kinds of different analytes anti--the particulate conjugate of the gold of analyte immunoglobulin G (dilution of described silver is called " mouse anti--analyte IgG solution II " later on).One of the people finger 82 that above-mentioned saliva sample is provided being withstood before described swab depressed for 10 seconds, processed swab can be cultivated one minute.Then, above-mentioned finger is lifted away from described swab, is pressed at once on the reaction medium of immunoassay apparatus, and be held in place for 30 seconds.Afterwards, make described finger be lifted away from this reaction medium, and, before recognition results, make this medium can cultivate two minutes.At this moment, the marking that central nucleus comprise that finger of that people that above-mentioned saliva sample is provided to zone 16, and also conversion zone 32 all develop the color to 37, shows the negative assay for all six kinds of analytes of existence.

Before being added to a new sterile swab, contain more a spot of Corvitin, tetrahydrocannabinol and ethanol to one and be added on this swab this point, repeat above-mentioned check program with described new sterile swab in-analyte IgG solution II anti-mouse.Finish after the check, immunoassay apparatus has shown the result shown in Fig. 2.Central nucleus comprise the marking of that finger of that people that above-mentioned saliva sample is provided to zone 16.Conversion zone 32 to 34 colour developings (negative findings), and conversion zone 35 to 37 is white (positive findingses).

Example 6

As in the example 5, prepare immunoassay apparatus.Below a tongue that will be verified a people who has or not six kinds of analytes, collect saliva sample with a sterile swab.When collecting this saliva sample, the mouse of 800 microlitres is resisted-analyte IgG solution I is placed in the sample tube with the dilute solution of a PCP.Make the swab contain saliva can soak the solution two minutes in this sample tube.Aliquot 400 microlitres of the potpourri that finally obtains are applied on the reaction medium, and make 20 seconds of potpourri aliquot that finally obtain that to cultivate this 400 microlitre.Then, join aliquot 400 microlitres and feasible can the discharge of solution A in the described reaction medium.After racking up solution A, be applied to that people's that above-mentioned saliva sample is provided a finger with the band gold label solution of 15 microlitres.The reaction medium that makes this finger withstand the said determination device is at once slowly depressed, and is held in place for 5 seconds.This finger is carefully rolled after described reaction medium, cultivating for 15 seconds before making this medium to join this reaction medium in the aliquot of the solution A of other 400 microlitres.Finish after the said procedure, conversion zone is white fully, shows the positive assay for PCP.Central nucleus comprise a marking of that finger of that people that above-mentioned saliva sample is provided to the zone.

Example 7

Method according to example 1 prepares immunoassay apparatus.Reaction medium comprises a nylon membrane.Conversion zone comprises some granules of polystyrene things, and these granules of polystyrene things are coated with the human serum albumin-cocaine that is embedded in the above-mentioned film.Central nucleus comprise some granules of polystyrene things to the zone, and these granules of polystyrene things resist-the human immunoglobulin coating with goat, are embedded in the above-mentioned film.

With the broken finger that will be verified the people who has or not cocaine in the blood of an acupuncture, so that obtain a droplet blood.Spread this droplet blood out with an aseptic spatula, so that on finger tip, form a blood film.Make the finger tip of this blood of having spread out withstand reaction medium and depress and be held in place about 10 seconds.Be lifted away from after this finger, handle described reaction medium, thus, get on except that the redness of blood from this reaction medium with the dilute solution of house detergent.After the redness of removing blood, with alkaline phosphate carry out the mouse of conjugation anti--dilute solution of cocaine immunoglobulin G handles above-mentioned reaction medium, and make and can cultivate this reaction medium two minutes.Then, order is with a kind of dilute solution of solution A, 5-bromo-4-chloro-3-indyl phosphate and nitroblue tetrazolium (BCIP/NBT), handle this reaction medium with solution A again.Make this reaction medium to handle and final solution A is cultivated a few minutes between handling at described BCIP/NBT.After finishing last program, central nucleus comprise a marking of that finger of that people that above-mentioned blood sample is provided to the zone, and the conversion zone colour developing, show the negative assay for cocaine.

Example 8

As in the example 7, prepare immunoassay apparatus.Mouse 500 microlitres, in the salt solution of the tween-20 that contains 0.1% bovine serum albumin(BSA) (BSA) and 0.05% of phosphate-buffered anti--aliquot of the dilute solution of cocaine immunoglobulin G (be called later on " mouse is anti--cocaine IgG solution) is placed in the small sample test tube.A solution that contains more a spot of cocaine is joined in this sample tube.

With the broken finger that will in their blood, be verified the people who has or not cocaine of an acupuncture,, and two droplet of blood are joined in the potpourri in the sample tube so that obtain a droplet of blood.Making the potpourri that finally obtains can cultivate two minutes in this sample tube after, this potpourri is joined on the reaction medium of immunoassay apparatus.On this reaction medium, cultivated this potpourri one minute, then, clean this potpourri with solution A.Band gold label solution with 15 microlitres is smeared that people's that above-mentioned blood sample is provided a finger.5 seconds were depressed and be held in place to the reaction medium that makes this finger withstand the said determination device lentamente, and then, careful ground roll is from this reaction medium.Before joining the aliquot of the solution A of 400 other microlitres on this reaction medium, make and to cultivate 15 seconds of this reaction medium.Finish after the said procedure, conversion zone is white (for the positive check of cocaine) fully, and checks the marking that the zone comprises that finger of that people that above-mentioned blood sample is provided.

Example 9

Except immunoassay apparatus comprises 6 conversion zones, prepare the immunoassay apparatus (see figure 1) according to the method for example 1.Each conversion zone comprises some granules of polystyrene things, and these granules of polystyrene things apply with the cholesterol conjugate that is embedded in the difference amount in the film.Conversion zone 32 comprises a kind of cholesterol conjugate, and this cholesterol conjugate is when producing positive signal when a kind of biofluid more than or equal to 150 milligrams of/deciliter whole cholesterol levels contacts with having.Conversion zone 33 to 36 includes the cholesterol conjugate of moderate, and wherein 35 orders increase the amount of cholesterol conjugate from zone 32 to the zone, i.e. 180mg/dl, 240mg/dl, 280mg/dl.Above-mentioned reaction medium comprises that also two references corresponding to 30ml/dl and 65ml/dl high-density lipoprotein (HDL) check the zone.

With the broken finger that will in their blood, be verified the people of cholesterol amount of an acupuncture, so that obtain a droplet of blood.This droplet of blood is joined in the small sample test tube, this test tube contain 400 milliliters, the mouse in the salt solution of phosphate-buffered is anti--cholesterol immunoglobulin G and the mouse of collaurum conjugation be anti--dilute solution of cholesterol immunoglobulin G, described salt solution contains 0.1% bovine serum albumin(BSA) (BSA) and 0.05% Tween-20 (later above-mentioned dilute solution is called " mouse resists-cholesterol IgG solution ").After cultivating three minutes, the film of said mixture is spread out on that people's that above-mentioned blood sample is provided a finger tip with an aseptic spatula.The reaction medium that makes this finger tip withstand above-mentioned immunoassay apparatus is at once depressed lentamente.Keep this finger tip after 5 seconds in place, make this finger be lifted away from described reaction medium carefully.Cultivated this reaction medium one minute, and handled with the dilute solution of house detergent then, so that get on except that the redness of blood from this reaction medium.After finishing said procedure, check the marking that the zone comprises that finger of that people that above-mentioned blood sample is provided.Conversion zone 32 to 33 colour developings (for the feminine gender check of cholesterol), and conversion zone 34 and 35 is white (for positive checks of cholesterol).When the cholesterol of predetermined threshold value amount existed at least, each conversion zone provided a positive check for cholesterol.For each conversion zone, corresponding to the amount of the cholesterol conjugate that in that regional reaction medium, is absorbed, above-mentioned threshold value cholesterol concentration difference.

Example 10

Be the inventory that to determine a large amount of other medicines of whether existing with the present invention below.This is exemplary, is not intended that the invention be limited to this. The 6-blocker DiureticsAcebutolol acetazolamide alprenolol amiloride Atenolol bendroflumethiazide

Happy and some the relevant compound furosemides of 6-boldenone-17-α-propiolactone 5-(1-hydroxyl-2-(1-first 3-fourth amino-4-phenoxy group-5--3-phenylpropyl alcohol amino) ethyl) the salicylamide sulfamoylbenzoic acid Metoprolol chlorine mercury methoxy third urine nadolol chlorthalidone oxprenolol dichlorphenamidum inderal ethacrynic acid gains in depth of comprehension

Hydrochlorothiazide

Mercusal

Aldactone

Three aminopterin and the compound that some are relevant Stimulant The arcotic antalgesicDiethylpropion pacify you the pain amphetamine to phenalgin ethyl Normeperidine amphetaminil BUP 2; 4-diaminourea-5-benzene thiazole codeine Benzphetamine Hydrochloride pain caffeine Oextropropoxyphene norpseudoephedrine heroin chlorobenzoxamine paracodin clobenzorex 4 that looses, 4-diphenyl-6-piperidyl oneself-3-ketone isoprophenamine six hydrogen-1-first-4-phenyl nitrogen cover-4-carboxylic acid second

Ester cocaine dionin N-(1-diformazan carbamyl propyl group)-N-hydroxyl first is at propyl group crotonamide crotethamide Ah rice pain two Corvitin morphine ephedrine nalbuphine etafedrine pentazocine vanillic diethylamide demerol N-ethylo benzene second propylamine NIH-7519 euvitol Y-promedols and the relevant compound amfetyline desobesi Furfenorex mefenorex Corvitin Euspirol methylephedrine ritalin Novartrina Coraminum pemoline Liticon 3 of muttering; 4-diformazan-2-phenylmorpholine 3-first-2-phenylmorpholine α; Ionamin N-norephedrine α, α--diphenyl-2-piperazine shallow lake methyl alcohol 1-phenyl-2-pyrrolidinyl pentane CHP-depot 1-(4-tolyl)-2-(1-pyrrolidinyl)-1-pentanoneStrychnia and the compound that some are relevant Protein stimulatory synthesizes ketosteroid Psychedelic7 α; Oneself sends pyridine (PCP) FL ketamine Pro-viron 2,5-dimethoxy-4-Metamfetamine hydrogen methyltestosterone THC protobolin marjunana methylandrostenolone methyltestosterone 19-nortestosterone second promise ketone anavar Oxymesterone adroyd stanozolol 17-clausterone ergotic acid boldenone diacetayl amide chlorotestosterone Mescaline dehydrogenation methyltestosterone phenyl ringTestosterone and the compound that some are relevant Opiate Sedative/hypnoticThe peaceful pethidine methaqualone of heroin chloral hydrate morphine glutethimide methadone (Methandone) the codeine third oxygen sweet smell Barbiturates Benzodiazepine cover classThe stable amobarbital chlorine nitrogen cover quinalbarbitone librium phenobarbital of the amytal allylbarbital flurazepam neo-barb L0 that oxazepans

ALprazolanic Antipsychotic drug/antidepressants SolventChlorpromazine ethanol Trazodone methyl alcohol haloperole isopropyl alcohol chlorine piperazine oxygen cover diglycol carbonate lithium chloroform Imipramine Antalgesic Anabolic steroidAspirin testosterone paracetamol methyltestosterone brufen 19-nortestosterone diflunisal stanozolol phenylbutazone anavar

Protobolin

Chlorotestosterone

Pro-viron

Second promise ketone

Example 11

Following table has been established four kinds of typical, generation substrates water-fast product, that add lustre to, and described water-fast product can be used from the present invention with a kind of suitable enzyme conjugate one, rather than is used from the present invention with aforesaid colloid gold label one.Right labeling method is well-known to adopt some ketone/chromogens, and those of ordinary skills are easy to implement.

Produce the substrate that adds lustre to of water-fast product

Enzyme	Substrate	Abbreviation	Original color	Final color
Enzyme	Substrate	Abbreviation	Original color	Final color	Horseradish peroxidase alkaline phosphatase galactosidase	The diaminobenzidine AEC 4-chloro that diaminobenzidine strengthens with nickel-1-naphthols naphthols-AS-BI-phosphate/fast red TR naphthols-AS-MX-phosphate/fast red TR naphthols-AS-BI-phosphate/new fuchsin bromine chloro-indole based phosphates/nitroblue tetrazolium 5-bromo-4-chloro-3-indyl-B-d-galactolipin pyranoside naphthols-AS-BI-B-d-galactolipin pyranoside	DAB DAB/ nickel AEC ... NABP/F R NAMP/F R NABP/N F BCIP/ NBT BCIG NABG	The clarification of the clarification of the clarification of the clarification of the clarification of the clarification of the clarification of the clarification of the clarification of clarification	The blue look redness of the red blue red reddish violet of brown ash/black

Example 12

In this example, adopt the apparatus and method of example 7, except following these aspects, these aspects are that determinator comprises shown in the example 3, is used for the conversion zone of those analytes, with the surface that solution A is got reaction medium in advance wet, puncture the finger of checked object, so that produce the blood of q.s, these blood are coated on the finger tip of this checked object, and make applied blood dry on the finger of this object.Then, the finger of the dry blood of band is applied on the surface of determinator, and maintenance contact about 15 seconds with this device.After this finger was lifted away from, the dilute solution processing reaction medium with the reagent of clarifying thus, got on except that the redness of blood from this reaction medium.After the redness of having removed blood, handle above-mentioned reaction medium with the dilute solution of some conjugates that is attached to the collaurum on a kind of secondary antibodies, every kind of conjugate is specially in conjunction with those analytes of above-mentioned appointment.After cultivating about two minutes, with a finger tip about 10 microlitres, be not used for applying above-mentioned blood to a kind of collaurum coating that is specifically designed to the ligand of the ligand acceptor in checking the zone by conjugation.After taking away this finger tip, central nucleus comprise a marking of that finger of that people that above-mentioned blood sample is provided to the zone, and above-mentioned conversion zone colour developing shows the negative assay for described those analytes.

Example 13

In this example, adopt the apparatus and method of example 7, except following these aspects, these aspects are that determinator comprises shown in the example 3, is used for the conversion zone of those analytes, and the surface of reaction medium is dry, stabs out the finger of checked object, so that produce the blood of q.s, this blood is coated on the finger tip of this checked object, and applied blood (before having done) at once is applied to about 15 seconds kinds on the reaction medium.After being lifted away from this finger, make that reaction medium can dry (typically, about 1 to 2 minute), handle described reaction medium with clear solutions, so that remove red haemocyte, and with the above-mentioned reaction medium of solution-treated of the reagent of clarifying, thus, get on except that the redness of blood from this reaction medium.After the redness of having removed blood, use the dilute solution of some conjugates of the collaurum that is attached to a kind of secondary antibodies to handle described reaction medium, on the special combination of every kind of conjugate, state one of those analytes of appointment.Cultivate after two minutes, with about 10 microlitres, conjugation applies a finger tip that is not used for applying above-mentioned blood to the collaurum of a kind of ligand that is specifically designed to the ligand acceptor in checking the zone.After taking away this finger tip, central nucleus comprise a marking of that finger of that people that above-mentioned blood sample is provided to the zone, and the conversion zone colour developing shows the negative assay for described those analytes.

Example 14

Method according to example 1 prepares a kind of determinator that contains 5 conversion zones (zone 32 in the corresponding diagram 1 and 34 to 37).As in the example 3, conversion zone comprises some polystyrene latexs, and these latex scribble cocaine (32), PCP (34), amphetamine/Corvitin (35), tetrahydrocannabinol (36) and ethanol (37) respectively as analyte.But, in this example, zone 33 does not comprise any absorbed analyte or antibody, but checks the zone as a reference, and promptly one can keep the zone of white fully when test ending.Central nucleus prepare as in the example 1 zone (corresponding to " identity is checked the zone " in the zone among Fig. 1 16), except it comprises some granules of polystyrene things, and these granules of polystyrene things are coated with the goat anti-rabbit immunoglobulin G that is embedded in the film.

With the solution A processing reaction medium A of 500 microlitres, thus, this reaction medium of getting wet.Obtain blood sample on one's body from a checked object, so that check has or not five kinds of analytes in its blood.The part of this sample is coated on the finger tip of described checked object, and is applied to five conversion zones and with reference to checking on the zone.On above-mentioned reaction medium, keep-uped pressure about 10 seconds to 15 seconds.

Some, for example, aliquot 500 microlitres, dilution house detergent solution is added on the reaction medium, and makes and can discharge then.House detergent is handled any blood cell that has the redness in the above-mentioned reaction medium of dissolving, and the dark red color of blood is washed from this medium.Then, with the mouse in the TRIS buffering agent about 10ml, the collaurum conjugation anti--dilute solution of the potpourri of analyte immunoglobulin G handles five conversion zones and with reference to checking the zone, this TRIS buffering agent contains 1.0% BSA, 0.05% Tween-20 and 0.05% azide.

Then, with the rabbit in the TRIS buffering agent 15 microlitres, the collaurum conjugation anti--dilute solution of goat immunoglobulin G smears that people's that blood sample is provided a finger, this TRIS buffering agent contains 1.0% BSA, 0.05% tween-20 and 0.05% azide.Depress lentamente in the identity authentication zone that makes above-mentioned finger withstand reaction medium, and be held in place for 3 seconds.After described finger carefully is lifted away from above-mentioned reaction medium, makes and before being added on this reaction medium, can cultivate 15 seconds of this medium to the aliquot of the solution A of other 400 microlitres.

After finishing said procedure,

conversion zone

32 and 34 to 37 colour developings show the negative assay for those analytes.With reference to checking zone 33 is white fully, and identity is checked the marking that the zone comprises that finger of that people that above-mentioned blood sample is provided.

Although the present invention has been described, be not that plan is confined to these embodiment according to exemplary embodiments.Those of ordinary skills particularly according to aforementioned techniques, can make embodiment that some still just are included in the scope of the present invention, alternative, example and improvement.Therefore, following claims attempt to cover any alternative embodiment, example, improvement or the equivalent that can be included in by in the determined the spirit and scope of the present invention of these claims.

Claims

1. a mensuration verifying attachment of providing for oneself is used to detect one by the analyte with check sample that checked object provided of differentiating body part surface, and measures verifying attachment and comprises:

(a) shell;

(b) a kind of reaction medium that is arranged in described shell comprises that at least one conversion zone and one are to differentiate the zone of checking by the checked object of main part surface imprint;

(c) a kind ofly be arranged in described shell and contain a kind of medium that produces the reagent of signal;

(d) a kind of medium that is used for holding a kind of check sample that is positioned at described shell; Described medium (c) and (d) each can independently move between a primary importance and a second place; Separate each other in described primary importance and described reaction medium; And contact with described reaction medium on the transferable material of described second place ground.

3. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that described at least one conversion zone comprises a kind of reagent that is used for determining to have or not at a check sample analyte.

4. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 3 is characterized in that described analyte is a kind of medicine or drug metabolite.

5. a kind of mensuration pick-up unit of providing for oneself as claimed in claim 1 is characterized in that described conversion zone comprises a kind of medicine or drug metabolite.

6. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that described at least one conversion zone comprises the member that a kind of ligand/acceptor is right.

7. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 6 is characterized in that the right member of described a kind of ligand/acceptor is a right member of a kind of antigen/antibody.

8. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that described reaction medium is a kind of low-protein combination, hydrophilic polysulfone membrane.

9. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that the described member that the zone comprises that a kind of ligand/acceptor is right that checks.

10. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that described reaction medium comprises the member that a kind of ligand/acceptor is right, and this ligand/acceptor pair constitutes a kind of conjugate with a kind of human antibodies to certain medicine.

11. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that the reagent of described generation signal is attached on the right member of a kind of ligand/acceptor.

12. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1, the reagent that it is characterized in that described generation signal are a kind of right members of ligand/acceptor with colloid gold label.

13. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 9, the reagent that it is characterized in that described generation signal with constitute a kind of conjugate the described described member that to check a kind of ligand/acceptor in the zone right.

14. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1, it is characterized in that the described medium that is used to hold a kind of check sample comprise a kind of can with the testing reagent of a kind of analyte reaction of seeking.

15. a kind of mensuration pick-up unit of providing for oneself as claimed in claim 1, it is characterized in that the described medium that is used to hold a kind of check sample comprises the member that a kind of ligand/acceptor is right, this ligand/acceptor is to constituting a kind of conjugate with a kind of analyte of seeking.

16. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that the described medium that is used to hold a kind of check sample is installed in a cylindrical end.

17. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that the described medium that is used to hold a kind of check sample comprises a kind of water wettability absorbing film.

18. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 17 is characterized in that the described medium that is used to hold a kind of check sample comprises a kind of water wettability spongy material.

19. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 1 is characterized in that arriving the contact that occurs transferable material under the pressure of 15psi at 2psi.

20. a mensuration verifying attachment of providing for oneself is used to detect one by the analyte with check sample that checked object provided of differentiating body part surface, and measures verifying attachment and comprises:

The shell of one two parts;

Reaction medium in a kind of one first parts of the shell that is installed in described two parts comprises that at least one conversion zone and one are to differentiate the zone of checking by the checked object of main part surface imprint;

In a kind of one second parts that contain the shell that is installed in described two parts certain produces the medium of the reagent of signal; With

A kind of removable medium that is used to hold a kind of check sample, this check sample is positioned at the centre of described first and described second parts of described shell, and, when described first and second parts of described shell are in one first detent position of described shell, contact above-mentioned check sample and the transferable material of described reaction medium;

The described medium that contains certain reagent that produces signal when described first and second parts of described shell are in one first detent position, with the transferable material of described reaction medium contact.

21. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 20 is characterized in that described first and second parts of described shell link together by a hinge.

22. a kind of mensuration verifying attachment of providing for oneself as claimed in claim 20, it is characterized in that the described member that the zone comprises that ligand/acceptor is right that checks, and described at least one conversion zone comprises and is used for determining that there is an a kind of right member of a kind of ligand/acceptor of analyte in above-mentioned check sample, it is described that to contain a kind of medium that produces the reagent of signal be a right member of a kind of ligand/acceptor with colloid gold label, the described removable medium that is used to hold a kind of check sample is a kind of water wettability spongy material, this water wettability spongy material comprises the member that a kind of ligand/acceptor is right, this kind ligand/acceptor is to forming a kind of conjugate with a kind of analyte of seeking, and described water wettability spongy material is installed on the cylindrical end.

23. a determinator is used to detect one by the analyte with check sample that checked object provided of differentiating body part surface, and measures verifying attachment and comprises:

(a) shell;

(b) a kind of reaction medium that is arranged in described shell comprises that at least one conversion zone and one are to differentiate the zone of checking by the checked object of main part surface imprint; And

(c) a kind of medium that is used for holding a kind of check sample that is positioned at described shell, and this medium can move between a primary importance and a second place, separate each other in described primary importance and described reaction medium, and the described second place and the transferable material of described reaction medium contact.

24. a kind of determinator as claimed in claim 23 is characterized in that occurring during greater than atmospheric pressure at pressure the contact of transferable material.

25. a kind of determinator as claimed in claim 23 is characterized in that at pressure be the contact that transferable material appears during to 15psi in 2psi.

26. a determinator is used to detect one by the analyte with check sample that checked object provided of differentiating body part surface, and measures verifying attachment and comprises:

(a) shell;

(c) a kind of a kind of medium that produces the reagent of signal that is arranged in described shell that holds, and this medium can move between a primary importance and a second place, separate each other in described primary importance and described reaction medium, and the described second place and the transferable material of described reaction medium contact.

28. a mensuration verifying attachment is used to detect one by the analyte with check sample that checked object provided of differentiating body part surface, and measures verifying attachment and comprises:

(a) shell;

30. an analysis box that is used to carry out a kind of mensuration, it comprises:

(i) a kind of mensuration verifying attachment is used to detect one by the analyte with check sample that checked object provided of differentiating body part surface; This device comprises:

(a) shell;

(c) a kind of a kind of medium that produces the reagent of signal that is arranged in described shell that holds, this medium can move between a primary importance and a second place, separate each other in described primary importance and described reaction medium, and the described second place and the transferable material of described reaction medium contact; With

(ii) a kind of reagent that comprises the member that a kind of ligand/acceptor is right, this ligand/acceptor is to forming a kind of conjugate with a kind of analyte.

32. a determination and analysis box, it comprises:

(i) a kind of mensuration verifying attachment is used to detect one by the analyte with check sample that checked object provided of differentiating body part surface, and measures this device of verifying attachment and comprises:

(a) shell;

(b) a kind of reaction medium that is arranged in described shell comprises that at least one conversion zone and one are to differentiate the zone of checking by the checked object of main part surface imprint; With

(c) a kind of medium that is used for holding a kind of check sample that is positioned at described shell, and this medium can move between a primary importance and a second place, separate each other in described primary importance and described reaction medium, and the described second place and the transferable material of described reaction medium contact;

(ii) a kind of reagent that produces signal, this reagent can be right with a kind of ligand/acceptor member reaction.

34. one kind is used for determining to have the method that there is a kind of analyte in the checked object of differentiating body part surface at one, the method comprises:

(a) when a kind of medium that is used to hold and shifts a kind of check sample is in a primary importance, the described medium that is used to hold and shifts a kind of check sample is contacted with an a kind of part of check sample, described primary importance and a kind of reaction medium separate each other, and this reaction medium comprises that at least one conversion zone and one are to differentiate the zone of checking by the checked object of main part surface imprint;

(b) move the described medium that is used to hold and shifts a kind of check sample to a second place, this second place and the transferable material of described reaction medium contact; And

(c) move a kind of medium that contains a kind of reagent that produces signal to a second place from a primary importance, separate each other in described primary importance and described reaction medium, and the described second place and the transferable material of described reaction medium contact; And

(d) have or not above-mentioned analyte in definite check sample from above-mentioned reaction medium.

35. method of determining to exist a kind of analyte as claimed in claim 34, after it is characterized in that between above-mentioned reaction medium and the above-mentioned medium that contains a kind of reagent that produces signal, the contact of transferable material occurring, make the body part surface of a checked object place to such an extent that contact with the above-mentioned zone of checking.