EP0165072A2
Expired / Lapsed
EP
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Specific binding assay reagent medium, test kit and process
Assignee: Hitachi Chemical Diagnostics Inc
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Filing: 1985-06-14
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Granted: —
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Expires: 2005-06-14
Abstract
Abstract An attenuator is included in a reagent medium of a luminescent specific-binding assay to suppress undesirable extraneous light. In one such assay, an analyte in a sample is reacted with a specific binding partner attached to a solid surface, forming an immobilized pair at the surface. One member of the immobilized pair is then allowed to react with a specific binding partner previously conjugated with one component of a luminescent reaction system, and the remaining components are provided in the reagent medium. The resulting light emitted in the luminescent reaction is recorded on photographic film or othr photodetector as a measure of the presence and quantity of the analyte in the sample. The reagent medium containing the remaining luminescent reaction system components also includes an attenuator, preferably a dye, which absorbs light over a range of wavelengths including that of the light emitted in the luminescent reaction, the attenuator being present in an amount sufficient to suppress the extraneous light observed adjacent the solid surface in the absence of the attentuator. Reduction of such extraneous light sharpens the recorded luminescent image, thereby allowing a more precise analysis of the light intensity, and additionally reduces the occurrence of false positive images in the assay. In another embodiment, the attenuator preferentially absorb light emitted in remote portions of a reaction volume so that only light emitted from proximal portions reaches a measurement means. Reactions otherwise requiring a separation step may thereby be conducted homogeneously.
Inventors
- 👤 John F. Burd
- 👤 John W. Dyminski
- 👤 Vincent A. Marinkovich
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 16:32
CPC Classifications
G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/543; G01N33/531; G01N33/532; G01N33/533
Claims Analysis
Total Claims: 24
Claim 1
Independent Claim
76 words
A reagent medium for use in a specific binding assay, wherein light emitted by the reaction of the components of a luminescent reaction system is utilized as a measure of a reaction between two reactive substances, and wherein extraneous light is produced, the reagent medium comprising an attenuator which absorbs light at the wavelength of the light emitted by the luminescent reaction system, the attenuator being present in a concentration sufficient to suppress the extraneous light.
Claim 2
Dependent Claim
76 words
A reagent medium for use in a specific binding assay, wherein light emitted by the reaction of the components of a luminescent reaction system is utilized as a measure of a reaction between two reactive substances, and wherein extraneous light is produced, the reagent medium comprising an attenuator which absorbs light at the wavelength of the light emitted by the luminescent reaction system, the attenuator being present in a concentration sufficient to suppress the extraneous light.
Claim 3
Dependent Claim
21 words
The reagent medium of claim 1, wherein said reagent medium further comprises at least one component of the luminescent reaction system.
Claim 4
Dependent Claim
16 words
The reagent medium of claim 1, wherein the reactive substances are immobilized at a solid surface.
Claim 5
Dependent Claim
14 words
The reagent medium of claim 1, wherein said specific binding assay is an immunoassay.
Claim 6
Dependent Claim
13 words
The reagent medium of claim 1, wherein said reagent medium is a liquid.
Claim 7
Dependent Claim
14 words
The reagent medium of claim 1, wherein the emitted light is produced by chemiluminescence.
Claim 8
Dependent Claim
13 words
The reagent medium of claim 1, wherein the light is emitted by luminol.
Claim 9
Dependent Claim
12 words
The reagent medium of claim 1, wherein said attenuator is a dye.
Claim 10
Dependent Claim
17 words
The reagent medium of claim 1, wherein said attenuator is a mixture of red and yellow dyes.
Claim 11
Dependent Claim
51 words
A test kit comprising: means for conducting a luminescent specific binding assay, said means including at least one of the components of a luminescent reaction system; and a reagent medium, said reagent medium including an attenuator which absorbs light at the wavelength of the light emitted by the luminescent reaction system.
Claim 12
Dependent Claim
51 words
A test kit comprising: means for conducting a luminescent specific binding assay, said means including at least one of the components of a luminescent reaction system; and a reagent medium, said reagent medium including an attenuator which absorbs light at the wavelength of the light emitted by the luminescent reaction system.
Claim 13
Dependent Claim
14 words
The test kit of claim 10, wherein said means further includes a test chamber.
Claim 14
Dependent Claim
16 words
The test kit of claim 10, wherein said means further includes at least one reactive ligand.
Claim 15
Dependent Claim
24 words
The test kit of claim 10, wherein said attenuator is provided in a mixture with at least one component of the luminescent reaction system.
Claim 16
Dependent Claim
12 words
The test kit of claim 10, wherein said attenuator is a dye.
Claim 17
Dependent Claim
227 words
A process for conducting a specific binding assay for the presence of an analyte in a test solution using light emitted in a luminescent reaction system as a measure of the presence of the analyte, the luminescent reaction system requiring at least two components, comprising the steps of: providing a test chamber having at least one face transparent to light, the chamber having therein adjacent the transparent face a test surface having attached thereto a substance reactive with the analyte; introducing the test solution into the test chamber so that the test solution is contacted to the test surface, whereby the analyte, if present in the test solution, is bound to the reactive substance attached to the test surface to produce an immobilized analyte; removing the test solution from the test chamber and washing the interior of the test chamber; introducing into the test chamber a specific binding partner for the analyte, said specific bending partner being conjugated with one component of the luminescent reaction system; removing the unreacted conjugated binding partner from the test chamber and washing the interior of the test chamber; introducing into the test chamber a reagent medium containing the remaining components of the luminescent reaction system and an attenuator which absorbs light at the wavelength of the light emitted by the luminescent reaction system; and recording the intensity of the light emitted.
Claim 18
Dependent Claim
227 words
A process for conducting a specific binding assay for the presence of an analyte in a test solution using light emitted in a luminescent reaction system as a measure of the presence of the analyte, the luminescent reaction system requiring at least two components, comprising the steps of: providing a test chamber having at least one face transparent to light, the chamber having therein adjacent the transparent face a test surface having attached thereto a substance reactive with the analyte; introducing the test solution into the test chamber so that the test solution is contacted to the test surface, whereby the analyte, if present in the test solution, is bound to the reactive substance attached to the test surface to produce an immobilized analyte; removing the test solution from the test chamber and washing the interior of the test chamber; introducing into the test chamber a specific binding partner for the analyte, said specific bending partner being conjugated with one component of the luminescent reaction system; removing the unreacted conjugated binding partner from the test chamber and washing the interior of the test chamber; introducing into the test chamber a reagent medium containing the remaining components of the luminescent reaction system and an attenuator which absorbs light at the wavelength of the light emitted by the luminescent reaction system; and recording the intensity of the light emitted.
Claim 19
Dependent Claim
11 words
The process of claim 15, wherein the attenuator is a dye.
Claim 20
Dependent Claim
114 words
A process for conducting a luminescent specific binding assay in a test chamber for an analyte in a test solution, using light emitted by a luminescent reaction system and recorded by a measurement means, comprising the steps of: immobilizing the analyte at a designated measurement location within the test chamber; introducing to the analyte a specific binding partner for the analyte, said binding partner b eeing conjugated to one of the components of the luminescent reaction system; and BAD ORIGINAL introducing into the. test chamber a reagent medium containing the remaining components of the luminescent reaction system, and an attenuator for suppressing the light produced at a location other than the designated measurement location.
Claim 21
Dependent Claim
114 words
A process for conducting a luminescent specific binding assay in a test chamber for an analyte in a test solution, using light emitted by a luminescent reaction system and recorded by a measurement means, comprising the steps of: immobilizing the analyte at a designated measurement location within the test chamber; introducing to the analyte a specific binding partner for the analyte, said binding partner b eeing conjugated to one of the components of the luminescent reaction system; and BAD ORIGINAL introducing into the. test chamber a reagent medium containing the remaining components of the luminescent reaction system, and an attenuator for suppressing the light produced at a location other than the designated measurement location.
Claim 22
Dependent Claim
18 words
The process of claim 17, wherein the analyte is immobilized at a solid surface within the test chamber.
Claim 23
Dependent Claim
11 words
The process of claim 17, wherein the attenuator is a dye.
Claim 24
Dependent Claim
37 words
The process of claim 17, including, after said step of introducing to the analyte and before said step of introducing into the test chamber, the further step of: removing the conjugated binding partner from the test chamber.
References Cited (Backward Citations)
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