Barrier-controlled assay device

A chromatographic assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a first opposable component including a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner for the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium;
• (ii) at least one absorber in operable contact with at least one of the first and second ends of the chromatographic medium;
• (iii) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (iv) an applicator layered on top of the barrier, the applicator containing a labeled specific binding partner to the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator, the applicator being positioned such that the barrier is located between the applicator and the chromatographic medium and such that an aqueous liquid applied to the applicator is drawn through the aperture after resolubilizing the labeled specific binding partner and then is drawn through the chromatographic medium by the at least one absorber so that the analyte and the labeled specific binding partner can form a ternary complex at the detection zone on the chromatographic medium;
• and
• (b) a second opposable component including a sample preparation zone;
• wherein the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in the sample preparation zone being in contact with the applicator so that sample in the sample preparation zone is applied to the applicator for resolubilization of the resolubilizable labeled specific binding partner and chromatography through the chromatographic medium.

A chromatographic assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium;
• (ii) at least one absorber in operable contact with at least one of the first and second ends of the chromatographic medium;
• and
• (iii) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (b) a second opposable component including an applicator for application of a sample thereto, the applicator containing a labeled specific binding partner to the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator;
• wherein the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in the applicator being in operable contact with the barrier so that the sample and the resolubilized labeled specific binding partner in the applicator are applied through the aperture in the barrier to the chromatographic medium for chromatography through the chromatographic medium.

An assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium;
• (ii) at least one absorber in operable contact with at least one of the first and second ends of the chromatographic medium;
• (iii) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (iv) a filter for removing particulates layered on top of the barrier;
• the chromatographic medium, the barrier, and the filter being positioned so that the barrier is located between the filter and the chromatographic medium in a fluid flow path transverse to the direction of flow along the chromatographic medium;
• and
• (b) a second opposable component including an applicator for application of a sample thereto, the applicator containing a labeled specific binding partner to the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator;
• wherein the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in the applicator being in operable contact with the filter so that the sample and the resolubilized labeled specific binding partner in the applicator are applied to the filter and then through the aperture in the barrier to the chromatographic medium for chromatography through the chromatographic medium.

An assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first and the second ends of the chromatographic medium;
• (ii) at least one absorber in operable contact with at least one of the first and second ends of the chromatographic medium;
• (iii) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• (iv) an applicator layered on top of the barrier, the applicator containing a labeled specific binding partner for the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator;
• and
• (v) a filter for removing particulates layered on top of the applicator, the chromatographic medium, the barrier, the applicator, and the filter being positioned so that the applicator is located between the filter and the chromatographic medium in a fluid flow path transverse to the direction of flow along the chromatographic medium;
• and
• (b) a second opposable component including a sample preparation zone;
• where the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in the sample preparation zone being in operable contact with the filter so that sample is applied to the filter, and a solution of filtered sample and labeled specific binding partner is then applied to the chromatographic medium through the aperture in the barrier.

An assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first end and the second end of the chromatographic medium;
• (ii) at least one absorber in operable contact with at least one of the first and second ends of the chromatographic medium;
• (iii) a first substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the first barrier at least partially blocking application of liquid to the chromatographic medium;
• (iv) an applicator layered on top of the first barrier, the applicator containing a labeled specific binding partner to the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator;
• (v) a second substantially fluid-impermeable barrier layered on top of a first portion of the applicator centrally located on the applicator;
• and
• (vi) a distribution membrane layered on top of the second barrier and in contact with portions of the applicator not supporting the second barrier so that fluid can flow from the distribution membrane to the applicator around the second barrier;
• and
• (b) a second opposable component including a receptacle for a swab that can contain a test sample;
• wherein the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in applying a sample to the distribution membrane and then to the chromatographic medium.

An assay device for performing a competitive immunoassay for detection and/or determination of an analyte in a test sample, comprising:

• (a) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a secondary specific binding partner immobilized thereon in a detection zone between the first and second ends of the chromatographic medium, the secondary specific binding partner capable of binding a member of a specific binding pair that lacks affinity for the analyte;
• (b) at least one absorber in operable contact with at least one of the first and second ends of the chromatographic medium;
• (c) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium, the barrier having at least one aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (d) an affinity membrane layered on top of the barrier, the affinity membrane containing a specific binding partner to the analyte immobilized thereto;
• the chromatographic medium, absorber, barrier, aperture, and affinity membrane being configured such a liquid applied to the affinity membrane is drawn through the chromatographic medium by the at least one absorber so that a labeled analyte analogue comprising analyte bound to a member of a specific binding pair lacking affinity for the analyte and bindable by the secondary specific binding partner, the analyte analog being labeled with a detectable label, binds at the detection zone on the chromatographic medium if analyte is present in the sample.

An assay device for performing a competitive immunoassay for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a secondary specific binding partner immobilized thereon in a detection zone between the first end and second end of the chromatographic medium, the secondary specific binding partner capable of binding a member of a specific binding pair that lacks affinity for the analyte, the secondary specific binding partner being an antibody specific for a species, class, or subclass of antibody that does not bind the analyte;
• (ii) a first absorber in operable contact with the first end of the chromatographic medium;
• (iii) a second absorber in operable contact with the second end of the chromatographic medium;
• (iv) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture therethrough for application of liquid to the chromatographic medium, the aperture being substantially smaller in area than the barrier so that liquid can enter the chormatographic medium only through the aperture;
• the chromatographic medium, absorbers, barrier and aperture being configured so that the detection zone is located between the aperture and the first end of the chromatographic medium;
• and
• (v) an affinity membrane layered on top of the barrier, the affinity membrane containing a specific binding partner to the analyte immobilized thereto;
• and
• (b) a second opposable component including an applicator, the applicator containing an analyte analogue in a form that can be resolubilized by the addition of an aqueous liquid to the applicator, the analyte analogue comprising analyte covalently bound to a member of a specific binding pair lacking affinity for the analyte and bindable by the secondary specific binding partner, the analyte analogue being labeled with a detectable label;
• wherein the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in the applicator being in operable contact with the affinity membrane.

An assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end and a second end and first and second surfaces, and having immobilized thereon in separate discrete and non-overlapping zones between the first and second ends of the chromatographic medium:
• (A) a detection zone with immobilized specific binding partner to the analyte;
• and
• (B) a control zone of analyte or analogue thereof;
• the detection zone being located between a point removed from the ends of the chromatographic medium and the second end of the chromatographic medium and the control zone being located between a point removed from the ends of the chromatographic medium and the second end of the chromatographic medium;
• (ii) an absorber in operable contact with the first end of the chromatographic medium;
• (iii) a first substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having a first application aperture substantially smaller than the area of the chromatographic medium and located at a point removed from the ends of the chromatographic medium, the first barrier at least partially blocking application of liquid to the chromatographic medium;
• (iv) an applicator layered on top of the first barrier, the applicator containing a labeled specific binding partner to the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator, the applicator positioned to apply liquid to the chromatographic medium via the first application aperture;
• and
• (v) a second substantially fluid-impermeable barrier layered on top of the applicator and positioned to at least partially block application of liquid to the applicator and to allow application of liquid to the applicator through a second application aperture substantially smaller than the area of the chromatographic medium and located closer to the second end of the chromatographic medium then to the first end of the chromatographic medium;
• and
• (b) a second opposable component including a sample preparation zone;
• wherein the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in the sample preparation zone being in contact with the second barrier so that the sample in the sample preparation zone is applied to the applicator through the second application aperture and traverses substantially the entire length of the applicator in order to apply the sample and the resolubilized labeled specific binding partner to the chromatographic medium through the first application aperture.

An assay device for detection and/or determination of a hydrophobic analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a nylon chromatographic medium having a first end, a second end, and first and second surfaces, the chromatographic medium having an affinity for a hydrophobic analyte sufficient to immobilize the analyte from an aqueous solution containing the analyte on the chromatographic medium;
• (ii) an applicator in operable contact with the first end of the chromatographic medium, the applicator containing a labeled specific binding partner to the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator;
• (iii) an absorber in operable contact with the second end of the chromatographic medium;
• and
• (iv) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (b) a second opposable component including a sample preparation zone;
• wherein the first and second opposable components are configured so that bringing the first and second opposable components into opposition results in the sample preparation zone being in contact with the barrier so that the sample in the sample preparation zone is applied to the chromatographic medium and so that analyte in a test sample is immobilized on the chromatographic medium in at least the vicinity of the aperture.

An assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner for the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium;
• (ii) a sample preparation zone in operable contact with the first end of the chromatographic medium;
• (iii) a first absorber in operable contact with the second end of the chromatographic medium;
• and
• (iv) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (b) a second opposable component including:
• (i) an applicator containing a labeled specific binding partner to the analyte in a form that can be resolubilized by the addition of an aqueous liquid to the applicator, the applicator positioned such that when the first and second opposable components are brought into opposition, the applicator is in contact with the barrier so that the resolubilized labeled specific binding partner is applied through the aperture in the barrier to the chromatographic medium;
• and
• (ii) second and third absorbers positioned such that when the first and second opposable components are brought into opposition, the second absorber and third absorber are in operable contact with portions of the chromatographic medium so that the second and third absorbers remove fluid from the chromatographic medium.

An assay device for detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first end and the second end of the chromatographic medium;
• (ii) an absorber in operable contact with the second end of the chromatographic medium;
• (iii) a conductor in operable contact with the first end of the chromatographic medium;
• (iv) a conjugate zone containing a specific binding partner to the analyte labeled with a gold sol in a form that can be resolubilized by the addition of an aqueous liquid to the conjugate zone, the conjugate zone being in direct contact with the conductor;
• (v) a sample preparation zone, the sample preparation zone being in direct contact with the conjugate zone, the conductor, the conjugate zone, and the sample preparation zone being positioned so that the conjugate zone bridges the sample preparation zone and the conductor;
• and
• (vi) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (b) a second opposable component including:
• (i) an applicator containing, in a form that can be resolubilized by the addition of an aqueous liquid to the applicator:
• (1) a soluble silver salt and (2) a reducing agent, the applicator positioned such that when the first and second opposable components are brought into opposition, the applicator is in contact with the barrier so that the resolubilized silver salt and the reducing agent are applied through the aperture in the barrier to the chromatographic medium;
• and
• (ii) second and third absorbers positioned such that when the first and second opposable components are brought into opposition, the second absorber and third absorber are in operable contact with portions of the chromatographic medium so that the second and third absorbers remove fluid from the chromatographic medium.

An assay device for the detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium;
• (ii) an absorber in operable contact with the second end of the chromatographic medium;
• (iii) a conductor in operable contact with the first end of the chromatographic medium;
• (iv) a conjugate zone containing a specific binding partner to the analyte labeled with a gold sol in a form that can be resolubilized by the addition of an aqueous liquid to the conjugate zone, the conjugate zone being in direct contact with the conductor;
• (v) a sample preparation zone, the sample preparation zone being in direct contact with the conjugate zone, the conductor, conjugate zone and sample preparation zone being positioned so that the conjugate zone bridges the sample preparation zone and the conductor;
• and
• (vi) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• and
• (b) the second opposable component including:
• (i) an applicator including two sectors:
• (A) a first sector for receiving an aqueous liquid;
• and
• (B) a second sector containing, in a form that can be resolubilized by the addition of an aqueous liquid to the second sector of the applicator:
• (1) a soluble silver salt and (2) a reducing agent, the applicator positioned so that when the first and second opposable components are brought into opposition, the first sector of the applicator is in direct contact with the aperture in the barrier and the second sector of the applicator is in indirect contact with the aperture so that the aqueous liquid is applied first to the chromatographic medium to provide a wash, followed by the aqueous silver salt and the reducing agent;
• and
• (ii) second and third absorbers positioned such that when the first and second opposable components are brought into opposition, the second absorber is in operable contact with a portion of the chromatographic medium between the barrier and the first end of the chromatographic medium and the third absorber is in operable contact with a portion of the chromatographic medium between the barrier and the second end of the chromatographic medium so that the second and third absorbers remove fluid from the chromatographic medium.

An assay device for the detection and/or determination of an analyte in a test sample comprising:

• (a) a panel including:
• (i) a first opposable component including:
• (A) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first end and the second end of the chromatographic medium;
• (B) a conjugate zone containing a specific binding partner to the analyte labeled with a gold sol in a form that can be resolubilized by the addition of an aqueous liquid to the conjugate zone, the conjugate zone being in operable contact with the first end of the chromatographic medium;
• (C) a conductor in operable contact with the conjugate zone, the conjugate zone bridging the first end of the chromatographic medium and the conductor;
• and
• (D) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• (ii) a second opposable component including:
• (A) a first receptacle for a swab which may contain a test sample;
• (B) a well for addition of at least one extraction reagent to the swab;
• and
• (C) a first absorber separated from the first receptacle and the well;
• and
• (iii) a second receptacle for holding a protrusion of an insert to align the insert with the first and second opposable components;
• the first and second opposable components of the panel being configured so that, when they are brought into opposition, the first absorber is brought into contact with a portion of the chromatographic medium, and the first receptacle is brought into contact with the conductor;
• and
• (b) an insert including:
• (i) an applicator containing, in a form that can be resolubilized by the addition of an aqueous liquid to the applicator:
• (1) a soluble silver salt and (2) a reducing agent;
• (ii) a second absorber;
• (iii) a third absorber;
• and
• (iv) a protrusion for insertion into the second receptacle of the panel;
• the insert configured so that when the protrusion is inserted into the second receptacle of the panel, the applicator is in operable contact with the aperture to apply the contents of the applicator to the chromatographic medium through the aperture, and the second and third absorbers are each in operable contact with a portion of the chromatographic medium to withdraw fluid from the chromatographic medium.

An assay device for the detection and/or determination of an analyte in a test sample comprising:

• (a) a panel including:
• (i) a first opposable component including:
• (A) a chromatographic medium having a first end, a second end, and first and second surfaces and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium and substantially smaller than the area of the chromatographic medium;
• (B) a conductor in operable contact with the first end of the chromatographic medium;
• and
• (C) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• (ii) a second opposable component including a first absorber, the first and second opposable components being configured so that when they are brought into opposition the first absorber is brought into operable contact with a portion of the chromatographic medium so that the first absorber contacts the chromatographic medium on the opposite side of the detection zone from the conductor;
• and
• (iii) a receptacle for holding a protrusion of an insert to align the insert with the first and second opposable components;
• and
• (b) an insert including:
• (i) an applicator containing, in a form that can be resolubilized by the addition of an aqueous liquid to the applicator, a substrate for an enzyme label that is bound to a specific binding partner to the analyte, the enzyme label producing an insoluble detectable product by catalysis of a reaction involving the substrate;
• (ii) a second absorber;
• (iii) a third absorber;
• and
• (iv) a protrusion for insertion into the receptacle of the panel;
• the insert configured so that when the protrusion is inserted into the receptacle of the panel, the applicator is in operable contact with the aperture to apply the contents of the applicator to the chromatographic medium through the aperture, and the second and third absorbers are each in operable contact with a portion of the chromatographic medium to withdraw fluid from the chromatographic medium.

An assay device for the detection and/or determination of an analyte in a test sample comprising:

• (a) a panel including:
• (i) a first opposable component including:
• (A) a chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium;
• (B) a conjugate zone containing a specific binding partner to the analyte labeled with an enzyme in a form that can be resolubilized by the addition of an aqueous liquid to the conjugate zone, the conjugate zone being in contact with the first end of the chromatographic medium;
• (C) a conductor in direct contact with the conjugate zone, the conjugate zone bridging the first end of the chromatographic medium and the conductor;
• and
• (D) a substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having an aperture for application of liquid to the chromatographic medium, the barrier at least partially blocking application of liquid to the chromatographic medium;
• (ii) a second opposable component including:
• (A) a first receptacle for a swab containing a test sample;
• (B) a well for addition of at least one extraction reagent to the swab;
• and
• (C) a first absorber separated from the first receptacle and the well;
• and
• (iii) a second receptacle for holding a protrusion of an insert to align the insert with the first and second opposable components;
• the first and second opposable components being configured so that when they are brought into opposition, the first absorber is brought into contact with a portion of the chromatographic medium so that the first absorber contacts the chromatographic medium on the opposite side of the detection zone from the conductor and the first receptacle is brought into contact with the conductor;
• and
• (b) an insert including:
• (i) an applicator containing, in a form that can be resolubilized by the addition of an aqueous liquid to the applicator, a substrate for the enzyme label bound to the specific binding partner to the analyte in the conjugate zone, the enzyme label producing a insoluble detectable product by catalysis of a reaction involving the substrate;
• (ii) a second absorber;
• (iii) a third absorber;
• and
• (iv) a protrusion for insertion into the second receptacle of the panel;
• the protrusion configured such that when the protrusion is inserted into the second receptacle of the panel, the applicator is in contact with the aperture to apply the contents of the applicator to the chromatographic medium through the aperture, and the second and third absorbers are each in contact with a portion of the chromatographic medium to withdraw fluid from the chromatographic medium.

An assay device for the detection and/or determination of an analyte in a test sample comprising:

• (a) a first opposable component including:
• (i) the chromatographic medium having a first end, a second end, and first and second surfaces, and having a specific binding partner to the analyte immobilized thereon in a detection zone between the first and second ends of the chromatographic medium;
• (ii) at least one absorber in operable contact with the one of the ends of the chromatographic medium;
• (iii) a first substantially fluid-impermeable barrier layered on top of the first surface of the chromatographic medium and having a first aperture therethrough for application of liquid to the chromatographic medium;
• (iv) a distribution membrane adjacent to the first substantially fluid-impermeable barrier to direct fluid to the first aperture in the first substantially fluid-impermeable barrier, the distribution membrane having a central portion and a portion surrounding the central portion;
• (v) a second substantially fluid-impermeable barrier layered on top of the central portion of the distribution membrane;
• (vi) an applicator containing a labeled specific binding partner to the analyte in resolubilizable form and layered on top of the second substantially fluid impermeable barrier and extending to make operable contact with the portion surrounding the central portion of the distribution membrane so that fluid can flow from the applicator around the second barrier to the portion of the distribution membrane surrounding the central portion;
• and
• (vii) a surface barrier containing a second aperture therethrough for application of liquid to the applicator;
• and
• (b) a second opposable component including a receptacle for a swab containing a test sample.