One-step enzymatic and amine detection technique
First Claim
1. A diagnostic kit for detecting an amine, enzyme, or an enzyme inhibitor within a test sample, the kit comprising:
- a plurality of reactive complexes that each comprises a substrate joined to a reporter and a separation species, said substrate being cleavable by an enzyme to release said reporter; and
a chromatographic medium that defines a first enzyme detection zone within which an enzyme detection signal is capable of being generated, wherein the presence or quantity of an enzyme, or an inhibitor thereof, is determinable from said enzyme detection signal, said chromatographic medium further defining an amine detection zone within which is contained a chemichromic dye, said chemichromic dye being capable of undergoing a color change in the presence of an amine, wherein the presence or quantity of an amine is determinable from said color change.
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Abstract
A technique for detecting the presence or quantity of an enzyme (or enzyme inhibitor) and/or an amine within a test sample is provided. For example, in one embodiment, a diagnostic test kit is employed that utilizes reactive complexes that each includes a substrate joined (e.g., covalently bonded, physically adsorbed, etc.) to a reporter and a separation species. Upon contacting the reactive complexes, enzymes may cleave the substrate and release the reporter. Moreover, the test kit may also employ a chemichromic dye, i.e., a dye that exhibits a detectable color change upon chemical reaction with one or more functional groups, such as amino groups. The signal generated (directly or indirectly) by the reporter and chemichromic dye may then be used to indicate the presence or quantity of an enzyme (or enzyme inhibitor) and amine, respectively, within the test sample.
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Citations
33 Claims
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1. A diagnostic kit for detecting an amine, enzyme, or an enzyme inhibitor within a test sample, the kit comprising:
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a plurality of reactive complexes that each comprises a substrate joined to a reporter and a separation species, said substrate being cleavable by an enzyme to release said reporter; and
a chromatographic medium that defines a first enzyme detection zone within which an enzyme detection signal is capable of being generated, wherein the presence or quantity of an enzyme, or an inhibitor thereof, is determinable from said enzyme detection signal, said chromatographic medium further defining an amine detection zone within which is contained a chemichromic dye, said chemichromic dye being capable of undergoing a color change in the presence of an amine, wherein the presence or quantity of an amine is determinable from said color change. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18)
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19. A diagnostic kit for detecting an amine or a hydrolytic enzyme within a test sample, the kit comprising:
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a plurality of reactive complexes that each comprises a substrate joined to a reporter and a specific binding member, said substrate being cleavable by a hydrolytic enzyme to release said reporter; and
a chromatographic medium that defines a first enzyme detection zone within which an enzyme detection signal is capable of being generated, wherein the presence or quantity of a hydrolytic enzyme is determinable from said enzyme detection signal, said chromatographic medium further defining an amine detection zone positioned downstream from said first enzyme detection zone, wherein a chemichromic dye is contained within said amine detection zone, said chemichromic dye being capable of undergoing a color change in the presence of an amine, wherein the presence or quantity of an amine is determinable from said color change. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26)
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27. A method for detecting an amine, enzyme, or enzyme inhibitor within a test sample, the method comprising:
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i) contacting the test sample with a chromatographic medium, said chromatographic medium defining an enzyme detection zone and an amine detection zone, wherein an enzyme detection signal is capable of being generated within said enzyme detection zone and an amine detection signal is capable of being generated within said amine detection zone;
ii) determining the presence or quantity of an enzyme or enzyme inhibitor from said enzyme detection signal; and
iii) determining the presence or quantity of an amine from said amine detection signal. - View Dependent Claims (28, 29, 30, 31, 32, 33)
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Specification