CA2127980A1 - Assay apparatus and method - Google Patents
Assay apparatus and methodInfo
- Publication number
- CA2127980A1 CA2127980A1 CA 2127980 CA2127980A CA2127980A1 CA 2127980 A1 CA2127980 A1 CA 2127980A1 CA 2127980 CA2127980 CA 2127980 CA 2127980 A CA2127980 A CA 2127980A CA 2127980 A1 CA2127980 A1 CA 2127980A1
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- Canada
- Prior art keywords
- assay
- species
- conjugate
- zone
- assayed
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
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Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54366—Apparatus specially adapted for solid-phase testing
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Hematology (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Clinical Laboratory Science (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
Assay apparatus comprises a first assay zone constituted by a foam pad (26) on a first solid support component (10) constituted by a plastics card having a channel (14) on one face thereof bordered by undercut walls (12) receiving a plastics slide (18) as a second solid support component having as a second assay zone a window (24). The support components fit together for relative sliding movement such that said zones are slidable with respect to one another from a first position in which said zones are separated from one another to a position in which said zones are brought into mutual contact by sliding motion. An antibody may be bound to the foam pad. An enzyme antibody conjugate may be unbound on the pad. An assay reagent which takes part in an observable reaction with the conjugate or a species to be assayed is present beneath window (24).
Description
WO 93/13856 PCl/GB93/~)0097 ASSAY APPARATUS AND METHOD ~12 ~8~ -The present invention relates to assay apparatus and a method of assay and has particular but not exclusive relevance to immuno-diagnostic assays.
Diagnostic tests based on the specificity of antibodies (e.g. monoclonal antibodies) and DNA technology are continually being developed and provide means for the identification of undesirable infections (bacterial, fungal or viral), or chemical entities, thus facilitating appropriate treatmen~s.
In addition to improvPments in the chemistry of diagnostic tests,~ there has been considerable development in the way in which the tests are form~tted. Initially, most diagnostic ~es~s were condu ted in the labora~ory due to the constraints imposed by the requirement for e~uipment, materials and ~rained operators. There have however been significant developments in format design which allow tests to be carried out in rela~ively short ~imescales, e.g. ten minutes or less, whilst requiring minimal input in terms of manual manipulations and operator skill.
Current imm~no-diagnostic tPsts rely on multi-step process2s requiring several manipulations and reasonably accurate timing o the steps. This is a major disadvantage in tests for use outside the laboratory such as in doctors surgeries, in the field or in the home. In these circumstan~es, ~est procedures may not be followed ~ery accurately by the end user. Although some test ki~s current~y available do not require many steps, they tend to be expensive as the technQlogy needed to achieve these requi~ements is inherently expensive. There is therefor~ a need for a simpler device reducing manufacturing costs whilst mai~taining simplicity of operation.
The present invention provides assay apparatus comprising a first assay zone on a first solid support component for capturing a species to be assayed thereon and a second assay zone on a second solid support component having an assay reagent thereon, means mounting said first and second solid support components together for relative sliding movement such that said zones are slidable with respect to one another from a first position in which said zones are separated from one another to a position in which said zones are brought into mutual contact by sliding motion.
~ he first and second support components may be different parts of a single:member or may be separate members mounted together. For instance, a single strip may be folded over to ` ~ring to end portions together and means may be provided to : hold the end portions face to face whilst allowing sliding movement thereof with respect to one another.
Preferably however, the first and second assay zones are provided on respective ones of a pair of plastics plate members constituting sald first and second solid support components.
: Preferably; one said plate member or one end region of such a strip has a pair of oppositely dirzcte~ channels ~: thereon in which edge regions of the other said plate membar are slidingly received to mount said plate members or said .
;~ strip re~ions in face:to f~ace sl~ding relationship.
: The species to be assayed may itself be such as to produce or catalyse~ an observable reaction with the assay ~: ~ reagent ~n the second assay zone. Otherwise, an assay reagent may be present unbound in said first reagent support zone which is a species capable of binding to th2 species to be assayed having :conjugated thereto a species capable of mediating an observable reaction in the presence of said assay ~: reagent of the second:assay zone.
: The observable reaction is preferably a colour forming reaction or a reac~ion giving rise to a sp~cies which is observable by fluorescence under UV light. Accordingly, one or both of the support componen~s is preferably transparent at least in its respective assay zone.
Diagnostic tests based on the specificity of antibodies (e.g. monoclonal antibodies) and DNA technology are continually being developed and provide means for the identification of undesirable infections (bacterial, fungal or viral), or chemical entities, thus facilitating appropriate treatmen~s.
In addition to improvPments in the chemistry of diagnostic tests,~ there has been considerable development in the way in which the tests are form~tted. Initially, most diagnostic ~es~s were condu ted in the labora~ory due to the constraints imposed by the requirement for e~uipment, materials and ~rained operators. There have however been significant developments in format design which allow tests to be carried out in rela~ively short ~imescales, e.g. ten minutes or less, whilst requiring minimal input in terms of manual manipulations and operator skill.
Current imm~no-diagnostic tPsts rely on multi-step process2s requiring several manipulations and reasonably accurate timing o the steps. This is a major disadvantage in tests for use outside the laboratory such as in doctors surgeries, in the field or in the home. In these circumstan~es, ~est procedures may not be followed ~ery accurately by the end user. Although some test ki~s current~y available do not require many steps, they tend to be expensive as the technQlogy needed to achieve these requi~ements is inherently expensive. There is therefor~ a need for a simpler device reducing manufacturing costs whilst mai~taining simplicity of operation.
The present invention provides assay apparatus comprising a first assay zone on a first solid support component for capturing a species to be assayed thereon and a second assay zone on a second solid support component having an assay reagent thereon, means mounting said first and second solid support components together for relative sliding movement such that said zones are slidable with respect to one another from a first position in which said zones are separated from one another to a position in which said zones are brought into mutual contact by sliding motion.
~ he first and second support components may be different parts of a single:member or may be separate members mounted together. For instance, a single strip may be folded over to ` ~ring to end portions together and means may be provided to : hold the end portions face to face whilst allowing sliding movement thereof with respect to one another.
Preferably however, the first and second assay zones are provided on respective ones of a pair of plastics plate members constituting sald first and second solid support components.
: Preferably; one said plate member or one end region of such a strip has a pair of oppositely dirzcte~ channels ~: thereon in which edge regions of the other said plate membar are slidingly received to mount said plate members or said .
;~ strip re~ions in face:to f~ace sl~ding relationship.
: The species to be assayed may itself be such as to produce or catalyse~ an observable reaction with the assay ~: ~ reagent ~n the second assay zone. Otherwise, an assay reagent may be present unbound in said first reagent support zone which is a species capable of binding to th2 species to be assayed having :conjugated thereto a species capable of mediating an observable reaction in the presence of said assay ~: reagent of the second:assay zone.
: The observable reaction is preferably a colour forming reaction or a reac~ion giving rise to a sp~cies which is observable by fluorescence under UV light. Accordingly, one or both of the support componen~s is preferably transparent at least in its respective assay zone.
The unbound assay reagent present in said first assay zone may be an antibody conjugate capable of binding to a species to be assayed. The term "antibody" in this context herein includes antibody fragments having selective affinity for antibody binding sites. It may be a conjugate between a nucleic acid or nucleic acid analogue for binding to a nucleic acid as ~he species to be assayed and a linker moiety such as biotin capable of binding to a conjugate between an enzyme for mediating such a reaction and a further linking moiety, e.g.
avidin or between said nucleic acid component and said enzyme component directly.
; ` - The assay reagent in said second assay zone may be a substrate for a co1our forming or other observable reaction mediated by said conjugate.
15The species to be assayed is to be captured on to the first assay zone so that its attachment is resistant to washin~ procedures during the assay. In some cases this may be achie~ed by direct physical capture or absorption. For inQtance, some bacteria or proteins will bind directly onto 20 a ~oughened plastics strip or mesh. A~ternatively and more i~
generally the first assay zone may have bound thereon an assay reagent which is capable of binding the species to be assayed.
This bound reagent may be an antibody, which in this context lncludes antibody molecules and also fragments thereof having selective bihding affinity for antibody binding sites, a nucleic acid or a nucleic acid ana~ogue, depending upon the nature of the species ~o be assayed.
Where the bound and/or the unbound assay reagent in the irst assay zone is an antibody or antibody conjug~te the antibody or antibody fragment is preferably a monoclonal antibody or fragment thereof.
Preferably, all of the reagents are present in dry form, the solvent ~usually water) needed for the assay reaction being derived from the addition of the sample.
35There may be an absorbent pad present in the firs~ assay zone for holding a quantity of liquid during the assay. This 7~ 4~
pad may for instance be of open cell foam or of absorbent paper such as filter paper or similar materials.
The arrangement may be such that the pad is compressed when the second solid support component is slid into position over the first assay zone.
The invention includes a method of assay comprising capturing a species to be assayed on a first assay zone provided on a solid support component, and (a) slidin~
thereover a second support component bearing a s~bstrate for an observable reaction mediated by said species to be assayed so as to c~ntact said substrate with said bound species to produce said observable reaction or (b) binding thereto a conjugate between an enzyme and a species capable of binding to the species to be assayed, washing to remove unbound coniugate and sliding thereover a second solid support component bearing a substrate for an observable reac~ion mediated by said conj~ga~e so as to contact said substrate with said bound conjugate to produce said observable reaction.
The inven~ion will be further described and illustrated by the following description of preferred embodiments with refPrence ~o the accompanying drawings in which:~
.
~igure 1 shows in perspective view an example of apparatus according to the invention in use in three stages of an assay procedure, and 25Figure 2 shows stages, including sample taking, involved in a specifi~ form of assay procedure described in Example 1 : below using the same apparatus according ~o the invention.
As shown in Figure 1, apparatus acoording to the invention comprises a irst solid support component 10 in the form of a plastics plate of about the size of a credit card having a pair of upstanding walls 12 bordering a central s~rip portion of lesser thickness 14. The faces of the walls 12 bordering the strip 14 are undercut as shown at 16..A second solid support co~ponen~ 18 comprises a plastics strip dimensioned to fit snugly between the walls 12 and having chamfered edges 20 which locate in the undercut shown at 16 -5~ 7~
to mount the support component 18 for sliding movement in the channel formed between the walls 12 on the support component 10. A fixst assay zone 22 is provided on the first solid support component 10 and a second assay zone 24 is provided on the second solid support component 18. At least the reyion of the said assay zone 24 on the second support component 18 is made transparent. A foam pad 26 is present in the first assay zone 22 securely adhered in place on the first support component 10.
10A substrate for an enzyme mediated colour forming . . reaction i~ present in dry form in the second assay zone 24.
: ; Whilst the above described features are sufficient to define an apparatus according to the invention in its broadest form, the apparatus illustrated in Figure 1 further comprises 15 unbound on the foam pad 26 an enzyme-an~ibody conjugata, the enzyme being sui~able to catalyse a co~our forming r~ac~ion with the substrate present on the second assay zone 24.
Also present in the foam pad 26 but bound thereon is a further an~ibody. The bound antibody and the antibody 20 component of the unbou~d antibody con;ugate may be the same o~ may be diff rent. Both are preferably monoclonal anti-bodies although polyclonal rea~ents are still suitable.
: :In use, a sample such as a liquid containing micro-~:organism cells is applied to the foam pad. The cells are 25 bound by the bound antibody and a "sandwich" is formed by ~binding to the bound cells of the antibody conjugate. The :apparatus may then be wa hed under running water, which may be tap water, to remove excess antibody conjugate from the sample area and the second support component 18 is then slid 30 over the first support component 10 to bring the first and second assay zones into position one over the other. The foam tfi~ter pad is thereby compressed against ~he undersidP of the second support component. Excess liquid is squeez~d ou~ from the foam pad by the leading edge of the second support 35 component passing over it and a good contact is ensured between the foam pad 26 and the second assay zone 24 which contains in dried form a substrate for the antibody conjugate.
This is resuspended by the residual liquid on the foam pad and the colour forming reaction takes place and is observable through the transparency of the second support component.
The following examples illustrate the use of the apparatus shown in Figure l.
EXAMYLE l - AssaY for EOcoli by Detection of 3-Glucuronidase There is a requirement in the wa~er industry for a convenient assay for detecting E. co~i in water. B-glucuronidase is an enzyme expressed by a limited number of '~ micro-organisms which is accepted in the water industry ~s being a 95~ positive test for E. coli. To exemplify the use of the apparatus described above, a sample of water is taken up into a syringe 27 and is discharged via a micro-organism trapping 0.2~m filte~ 28. The micro-organisms initi lly present in ~h~ liquid sample are now held in concentrated form in the li~uid in the dead space of the syringe/filter assembly. A defi~ed amount of a sui~able micro-organism ~20 growth medium is drawn up into ~he filter. The syringe/filtex :assembly is now placed with the syringe vertical and the lower face of the hou ing of the filter on a heating block by which the ilter is warmed to an incubating temperature. After a predetermined period of incubation to multiply the bacteria, ~:~25 the temperature of the heating block may be raised to a temperature suitable to produce lysis of the bacteria for a perlod of about one minute. Apparatus according to Figure l i~ :empIoyed having a polyclonal antibody to B-glucuronidase bound to the fsam pad 26. In this case no antibody conjugate is present on the foam pad as B-glucuronidase itself catalyses a colour forming reaction. The liquid content of ~he syringe/filter assembly may then ~e dischar~ed on to the first assay zone 22 of apparatus as shown in Figure l and may be left a~ room ~mperature for a period sufficient to enable binding of the B-glucuronidase from the micro-organism lysate :by the bound antibody on the first assay zon~, e.g. for about WO93/13856 PCT/GB93/~0097 ten minutes. The assay apparatus is optionally then washed and the second support component 18 is slid int~ position as shown ln step 3 in Figure 1. The colour forming reaction is allowed to take place and is observed.
Optionally, a series of syringe and filter assemblies are prepared at the outset of the procedure and these are incubated for differing periods of time, the 'ength of time necessary for incubation depending upon the l~vel of contamination present. Optionally, to enhance sensiti~i~y, the growth medium may contain a substrate suitable to induce expression of the B-glucuronidase by the micro-organisms to : .produce elevated levels of enzyme in the cultured cells, e.g.
an R-B-D-glucuronide. Also to enhance sensitivity the colour forming reaction may be replaced by a reac~ion giving rise to a W ~luorescent compound by appropriate selection of the enzyme substrate on the ~econd support component 18.
EXAMPLE 2 -_Sandwiçh AssaY for_B-Glucuronidase To enhance sensitivity the apparatus employ~d in Example 1 is modified by the inclusion of a con~ugate between an ant$
B-glucuronidase antibody and a -colour forming reaction catalysing enzyme in dried form unbound in said foam pad 26.
The proc~dure followed is as in Example 1 up to the end of the lysis s~ep. When the liquid conten~ of the syringe/filter assembly is dischar~ed on to the foam pad a sandwich is formed in which the conjuga~e and the bound antibody both bind to ~h~
B-glucuronidase. The foam ~ad i5 washed to remove unbound conjugate and the second support component is slid into position. The assay reagent thereon is selected to take part in a colour forming reaction in the presence of the bound coniugate.
XAMPLE 3 - Assay for ~-Glucuronidase Tra~ped Directl~
The procedure of Example is modiied in ~ha~ the bound antibody on the foam pad 26 is omitted and the fo~m material is selected such that it has a non-specific affinity for W093/13856 PCT/GB93/00097.
proteins including B-glucuronidase. The lysate as applied to the foam pad and proteins therein are bound to the foam.
Bound B-glucuronidase is further bound by the free antibody conjugate. The presence of bound conjugate is then detected by washing and developing as in Example 2. Alternatively, the antibody conjugate is also omitted and the trapped B-glucuronidase acts directly to catalyse the colour forming reaction as in Example 1.
Many modifications and variations of the met~ods and apparatus described above are within the scope of the ln.ention.
: :
, ':
avidin or between said nucleic acid component and said enzyme component directly.
; ` - The assay reagent in said second assay zone may be a substrate for a co1our forming or other observable reaction mediated by said conjugate.
15The species to be assayed is to be captured on to the first assay zone so that its attachment is resistant to washin~ procedures during the assay. In some cases this may be achie~ed by direct physical capture or absorption. For inQtance, some bacteria or proteins will bind directly onto 20 a ~oughened plastics strip or mesh. A~ternatively and more i~
generally the first assay zone may have bound thereon an assay reagent which is capable of binding the species to be assayed.
This bound reagent may be an antibody, which in this context lncludes antibody molecules and also fragments thereof having selective bihding affinity for antibody binding sites, a nucleic acid or a nucleic acid ana~ogue, depending upon the nature of the species ~o be assayed.
Where the bound and/or the unbound assay reagent in the irst assay zone is an antibody or antibody conjug~te the antibody or antibody fragment is preferably a monoclonal antibody or fragment thereof.
Preferably, all of the reagents are present in dry form, the solvent ~usually water) needed for the assay reaction being derived from the addition of the sample.
35There may be an absorbent pad present in the firs~ assay zone for holding a quantity of liquid during the assay. This 7~ 4~
pad may for instance be of open cell foam or of absorbent paper such as filter paper or similar materials.
The arrangement may be such that the pad is compressed when the second solid support component is slid into position over the first assay zone.
The invention includes a method of assay comprising capturing a species to be assayed on a first assay zone provided on a solid support component, and (a) slidin~
thereover a second support component bearing a s~bstrate for an observable reaction mediated by said species to be assayed so as to c~ntact said substrate with said bound species to produce said observable reaction or (b) binding thereto a conjugate between an enzyme and a species capable of binding to the species to be assayed, washing to remove unbound coniugate and sliding thereover a second solid support component bearing a substrate for an observable reac~ion mediated by said conj~ga~e so as to contact said substrate with said bound conjugate to produce said observable reaction.
The inven~ion will be further described and illustrated by the following description of preferred embodiments with refPrence ~o the accompanying drawings in which:~
.
~igure 1 shows in perspective view an example of apparatus according to the invention in use in three stages of an assay procedure, and 25Figure 2 shows stages, including sample taking, involved in a specifi~ form of assay procedure described in Example 1 : below using the same apparatus according ~o the invention.
As shown in Figure 1, apparatus acoording to the invention comprises a irst solid support component 10 in the form of a plastics plate of about the size of a credit card having a pair of upstanding walls 12 bordering a central s~rip portion of lesser thickness 14. The faces of the walls 12 bordering the strip 14 are undercut as shown at 16..A second solid support co~ponen~ 18 comprises a plastics strip dimensioned to fit snugly between the walls 12 and having chamfered edges 20 which locate in the undercut shown at 16 -5~ 7~
to mount the support component 18 for sliding movement in the channel formed between the walls 12 on the support component 10. A fixst assay zone 22 is provided on the first solid support component 10 and a second assay zone 24 is provided on the second solid support component 18. At least the reyion of the said assay zone 24 on the second support component 18 is made transparent. A foam pad 26 is present in the first assay zone 22 securely adhered in place on the first support component 10.
10A substrate for an enzyme mediated colour forming . . reaction i~ present in dry form in the second assay zone 24.
: ; Whilst the above described features are sufficient to define an apparatus according to the invention in its broadest form, the apparatus illustrated in Figure 1 further comprises 15 unbound on the foam pad 26 an enzyme-an~ibody conjugata, the enzyme being sui~able to catalyse a co~our forming r~ac~ion with the substrate present on the second assay zone 24.
Also present in the foam pad 26 but bound thereon is a further an~ibody. The bound antibody and the antibody 20 component of the unbou~d antibody con;ugate may be the same o~ may be diff rent. Both are preferably monoclonal anti-bodies although polyclonal rea~ents are still suitable.
: :In use, a sample such as a liquid containing micro-~:organism cells is applied to the foam pad. The cells are 25 bound by the bound antibody and a "sandwich" is formed by ~binding to the bound cells of the antibody conjugate. The :apparatus may then be wa hed under running water, which may be tap water, to remove excess antibody conjugate from the sample area and the second support component 18 is then slid 30 over the first support component 10 to bring the first and second assay zones into position one over the other. The foam tfi~ter pad is thereby compressed against ~he undersidP of the second support component. Excess liquid is squeez~d ou~ from the foam pad by the leading edge of the second support 35 component passing over it and a good contact is ensured between the foam pad 26 and the second assay zone 24 which contains in dried form a substrate for the antibody conjugate.
This is resuspended by the residual liquid on the foam pad and the colour forming reaction takes place and is observable through the transparency of the second support component.
The following examples illustrate the use of the apparatus shown in Figure l.
EXAMYLE l - AssaY for EOcoli by Detection of 3-Glucuronidase There is a requirement in the wa~er industry for a convenient assay for detecting E. co~i in water. B-glucuronidase is an enzyme expressed by a limited number of '~ micro-organisms which is accepted in the water industry ~s being a 95~ positive test for E. coli. To exemplify the use of the apparatus described above, a sample of water is taken up into a syringe 27 and is discharged via a micro-organism trapping 0.2~m filte~ 28. The micro-organisms initi lly present in ~h~ liquid sample are now held in concentrated form in the li~uid in the dead space of the syringe/filter assembly. A defi~ed amount of a sui~able micro-organism ~20 growth medium is drawn up into ~he filter. The syringe/filtex :assembly is now placed with the syringe vertical and the lower face of the hou ing of the filter on a heating block by which the ilter is warmed to an incubating temperature. After a predetermined period of incubation to multiply the bacteria, ~:~25 the temperature of the heating block may be raised to a temperature suitable to produce lysis of the bacteria for a perlod of about one minute. Apparatus according to Figure l i~ :empIoyed having a polyclonal antibody to B-glucuronidase bound to the fsam pad 26. In this case no antibody conjugate is present on the foam pad as B-glucuronidase itself catalyses a colour forming reaction. The liquid content of ~he syringe/filter assembly may then ~e dischar~ed on to the first assay zone 22 of apparatus as shown in Figure l and may be left a~ room ~mperature for a period sufficient to enable binding of the B-glucuronidase from the micro-organism lysate :by the bound antibody on the first assay zon~, e.g. for about WO93/13856 PCT/GB93/~0097 ten minutes. The assay apparatus is optionally then washed and the second support component 18 is slid int~ position as shown ln step 3 in Figure 1. The colour forming reaction is allowed to take place and is observed.
Optionally, a series of syringe and filter assemblies are prepared at the outset of the procedure and these are incubated for differing periods of time, the 'ength of time necessary for incubation depending upon the l~vel of contamination present. Optionally, to enhance sensiti~i~y, the growth medium may contain a substrate suitable to induce expression of the B-glucuronidase by the micro-organisms to : .produce elevated levels of enzyme in the cultured cells, e.g.
an R-B-D-glucuronide. Also to enhance sensitivity the colour forming reaction may be replaced by a reac~ion giving rise to a W ~luorescent compound by appropriate selection of the enzyme substrate on the ~econd support component 18.
EXAMPLE 2 -_Sandwiçh AssaY for_B-Glucuronidase To enhance sensitivity the apparatus employ~d in Example 1 is modified by the inclusion of a con~ugate between an ant$
B-glucuronidase antibody and a -colour forming reaction catalysing enzyme in dried form unbound in said foam pad 26.
The proc~dure followed is as in Example 1 up to the end of the lysis s~ep. When the liquid conten~ of the syringe/filter assembly is dischar~ed on to the foam pad a sandwich is formed in which the conjuga~e and the bound antibody both bind to ~h~
B-glucuronidase. The foam ~ad i5 washed to remove unbound conjugate and the second support component is slid into position. The assay reagent thereon is selected to take part in a colour forming reaction in the presence of the bound coniugate.
XAMPLE 3 - Assay for ~-Glucuronidase Tra~ped Directl~
The procedure of Example is modiied in ~ha~ the bound antibody on the foam pad 26 is omitted and the fo~m material is selected such that it has a non-specific affinity for W093/13856 PCT/GB93/00097.
proteins including B-glucuronidase. The lysate as applied to the foam pad and proteins therein are bound to the foam.
Bound B-glucuronidase is further bound by the free antibody conjugate. The presence of bound conjugate is then detected by washing and developing as in Example 2. Alternatively, the antibody conjugate is also omitted and the trapped B-glucuronidase acts directly to catalyse the colour forming reaction as in Example 1.
Many modifications and variations of the met~ods and apparatus described above are within the scope of the ln.ention.
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Claims (12)
1. Assay apparatus comprising a first assay zone on a first solid support component for capturing a species to be assayed thereon and a second assay zone on a second solid support component having an assay reagent thereon, means mounting said first and second solid support components together for relative sliding movement such that said zones are slidable with respect to one another from a first position in which said zones are separated from one another to a position in which said zones are brought into mutual contact by sliding motion.
2. Apparatus as claimed in Claim 1, wherein an assay reagent is present unbound in said first reagent support zone which is a species capable of binding to the species to be assayed having conjugated thereto a species capable of mediating an observable reaction in the presence of said assay reagent of the second assay zone.
3. Apparatus as claimed in Claim 2, wherein said unbound assay reagent present in said first assay zone is an antibody conjugate, which antibody conjugate is capable of binding to a species to be assayed and wherein said assay reagent in said second assay zone is a substrate for an observable reaction mediated by said antibody conjugate.
4. Apparatus as claimed in Claim 3, wherein said antibody conjugate is a monoclonal antibody conjugate.
5. Apparatus as claimed in Claim 1 or Claim 2, wherein said unbound assay reagent is a conjugate between an enzyme and a nucleic acid or nucleic acid analogue probe.
6. Apparatus as calmed in any preceding claim, wherein said first assay zone has bound thereon an assay reagent which is capable of binding a species to be assayed.
7. Apparatus as claimed in Claim 6, wherein said assay reagent bound in said first assay zone an antibody or antibody fragment, or a nucleic acid or nucleic acid analogue capture probe.
8. Apparatus as claimed in Claim 6, wherein said bound assay reagent is a monoclonal antibody.
9. Apparatus as claimed in any preceding claim, wherein said first and second assay zones are provided on respective ones of a pair of plastics plate members constituting said first and second solid support components.
10. Apparatus as claimed in Claim 9, wherein one said plate member has a pair of oppositely directed channels thereon in which edge regions of the other said plate member are slidingly received to mount said plate members in face to face sliding relationship.
11. A method of assay comprising capturing a species to be assayed on a first assay zone provided on a solid support component, and (a) sliding thereover a second support component bearing a substrate for an observable reaction mediated by said species to be assayed so as to contact said substrate with said bound conjugate to produce said observable reaction or (b) binding thereto a conjugate between an enzyme and a species capable of binding to the species to be assayed, washing to remove unbound conjugate and sliding thereover a second support component bearing a substrate for an observable reaction mediated by said conjugate so as to contact said substrate with said bound conjugate to produce said observable reaction.
12. A method as claimed in Claim 11, conducted using apparatus as claimed in any one of Claims 1 to 9.
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Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| GB929201089A GB9201089D0 (en) | 1992-01-18 | 1992-01-18 | A diagnostic article |
| GB9201089.1 | 1992-01-18 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2127980A1 true CA2127980A1 (en) | 1993-07-22 |
Family
ID=10708832
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2127980 Abandoned CA2127980A1 (en) | 1992-01-18 | 1993-01-15 | Assay apparatus and method |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP0621803A1 (en) |
| AU (1) | AU3359193A (en) |
| CA (1) | CA2127980A1 (en) |
| GB (1) | GB9201089D0 (en) |
| WO (1) | WO1993013856A1 (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5449794A (en) * | 1991-02-15 | 1995-09-12 | Jasmine Fockerman | Benzopyran phenol derivatives for use as antibacterial agents |
| KR19980703048A (en) * | 1995-03-20 | 1998-09-05 | 피터쥐.스트링거 | 5-substituted-3- (1,2,3,6-tetrahydropyridin-4-yl) - and 3- (piperidin-4-yl) -1H- indole: A novel 5-HT1F agonist |
| US5744096A (en) * | 1997-02-21 | 1998-04-28 | Cholestech Corporation | Automated immunoassay cassette |
| NL1017374C2 (en) * | 2001-02-15 | 2002-08-16 | Univ Delft Tech | Device for carrying out a reaction, as well as a method for carrying out a reaction in the device. |
| NL1019378C2 (en) * | 2001-11-16 | 2003-05-20 | Univ Delft Tech | Method for filling a well in a substrate. |
| EP1357383B1 (en) | 2002-04-09 | 2005-11-09 | Cholestech Corporation | High-density lipoprotein assay device and method |
| US20040101439A1 (en) * | 2002-11-21 | 2004-05-27 | Fusco Adam J | Biological and chemical reaction devices and methods of manufacture |
| US7343784B2 (en) * | 2005-06-28 | 2008-03-18 | Paavo Kinnunen | Method and device for forming a liquid—liquid interface, especially for surface tension measurement |
| WO2008086019A1 (en) | 2007-01-09 | 2008-07-17 | Cholestech Corporation | Device and method for measuring ldl-associated cholesterol |
| WO2010109445A1 (en) | 2009-03-23 | 2010-09-30 | Ramot At Tel-Aviv University Ltd. | Assay device and method |
| JP6039965B2 (en) * | 2012-08-22 | 2016-12-07 | 森永製菓株式会社 | Immunochromatography device |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3415361A (en) * | 1966-12-22 | 1968-12-10 | Miles Lab | Test device and container therefor |
| TW203120B (en) * | 1985-10-04 | 1993-04-01 | Abbott Lab | |
| US4978502A (en) * | 1987-01-05 | 1990-12-18 | Dole Associates, Inc. | Immunoassay or diagnostic device and method of manufacture |
| EP0299359A3 (en) * | 1987-07-16 | 1990-09-26 | Abbott Laboratories | Reagent delivery system for use in solid-phase analytical devices |
| US4859604A (en) * | 1987-08-27 | 1989-08-22 | Ampor, Inc. | Composition for stabilization of diagnostic reagents |
| US4877586A (en) * | 1988-07-27 | 1989-10-31 | Eastman Kodak Company | Sliding test device for assays |
| FR2637687B1 (en) * | 1988-10-11 | 1991-01-11 | Inst Textile De France | SINGLE USE DEVICE FOR BIOLOGICAL TESTS |
| NL9002708A (en) * | 1990-12-10 | 1992-07-01 | Livestock Control Holding | DISPOSABLE LIQUID TESTING DEVICE. |
-
1992
- 1992-01-18 GB GB929201089A patent/GB9201089D0/en active Pending
-
1993
- 1993-01-15 CA CA 2127980 patent/CA2127980A1/en not_active Abandoned
- 1993-01-15 AU AU33591/93A patent/AU3359193A/en not_active Abandoned
- 1993-01-15 WO PCT/GB1993/000097 patent/WO1993013856A1/en not_active Application Discontinuation
- 1993-01-15 EP EP93902397A patent/EP0621803A1/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| GB9201089D0 (en) | 1992-03-11 |
| WO1993013856A1 (en) | 1993-07-22 |
| AU3359193A (en) | 1993-08-03 |
| EP0621803A1 (en) | 1994-11-02 |
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Legal Events
| Date | Code | Title | Description |
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| FZDE | Dead |