MEANS AND METHODS FOR THE DETERMINATION OF (D)-2-HYDROXYGLUTARATE (D2HG) OR (D)-2-HYDROXYADIPIC ACID
First Claim
1. A method for detecting (D)-2-hydroxyglutarate or (D)-2-hydroxyadipic acid in a sample, the method comprising:
- a) contacting a sample with a reagent mixture, wherein said reagent mixture comprises;
(i) a solvent,(ii) a dye having an oxidized state and a reduced state, wherein the reduced state can be distinguished from the oxidized state and wherein the dye is initially present in the oxidized state,(iii) an electron transfer agent,(iv) a (D)-2-hydroxyglutarate dehydrogenase enzyme, and(v) a cofactor; and
b) detecting (D)-2-hydroxyglutarate or (D)-2-hydroxyadipic acid by measuring the production of the reduced state of the dye.
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Abstract
The present invention relates to a method for detecting (D)-2-hydroxyglutarate or (D)-2-hydroxyadipic acid in a sample, the method comprising the steps of: a) contacting a sample with a reagent mixture, wherein said reagent mixture comprises: (i) a solvent, (ii) a dye having an oxidized state and a reduced state, wherein the reduced state can be distinguished from the oxidized state and wherein the dye is initially present in the oxidized state, (iii) an electron transfer agent, (iv) a (D)-2-hydroxyglutarate dehydrogenase enzyme, and (v) a cofactor; and b) detecting (D)-2-hydroxyglutarate or (D)-2-hydroxyadipic acid by measuring the production of the reduced state of the dye. The invention further pertains to a method for diagnosing and/or monitoring a (D)-2-hydroxy-glutarate-associated disease in a subject. Encompassed by the invention is also a method for diagnosing a mutation in an isocitrate dehydrogenase (IDH) gene or in a (D)-2-hydroxyglutarate (D2HG) dehydrogenase enzyme gene in a subject. In addition, the invention provides for a kit comprising (i) a solvent, (ii) a dye having an oxidized state and a reduced state, wherein the reduced state can be distinguished from the oxidized state and wherein the dye is initially present in the oxidized state, (iii) an electron transfer agent, (iv) a (D)-2-hydroxyglutarate dehydrogenase enzyme, and (v) a cofactor.
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Citations
15 Claims
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1. A method for detecting (D)-2-hydroxyglutarate or (D)-2-hydroxyadipic acid in a sample, the method comprising:
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a) contacting a sample with a reagent mixture, wherein said reagent mixture comprises; (i) a solvent, (ii) a dye having an oxidized state and a reduced state, wherein the reduced state can be distinguished from the oxidized state and wherein the dye is initially present in the oxidized state, (iii) an electron transfer agent, (iv) a (D)-2-hydroxyglutarate dehydrogenase enzyme, and (v) a cofactor; and b) detecting (D)-2-hydroxyglutarate or (D)-2-hydroxyadipic acid by measuring the production of the reduced state of the dye. - View Dependent Claims (7, 8, 9, 10, 11, 12, 13, 14)
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2. A method for diagnosing and/or monitoring a (D)-2-hydroxyglutarate-associated disease in a subject, the method comprising:
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a) contacting a sample of said subject with a reagent mixture, wherein said reagent mixture comprises; (i) a solvent, (ii) a dye having an oxidized state and a reduced state, wherein the reduced state can be distinguished from the oxidized state and wherein the dye is initially present in the oxidized state, (iii) an electron transfer agent, (iv) a (D)-2-hydroxyglutarate dehydrogenase enzyme, and (v) a cofactor; and b) detecting (D)-2-hydroxyglutarate by measuring the production of the reduced state of the dye, thereby diagnosing and/or monitoring a (D)-2-hydroxyglutarate-associated disease in said subject. - View Dependent Claims (3, 4)
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5. A method for diagnosing a mutation in an isocitrate dehydrogenase (IDH) gene or in a (D)-2-hydroxyglutarate (D2HG) dehydrogenase gene in a subject, the method comprising:
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a) contacting a sample of said subject with a reagent mixture, wherein said reagent mixture comprises; (i) a solvent, (ii) a dye having an oxidized state and a reduced state, wherein the reduced state can be distinguished from the oxidized state and wherein the dye is initially present in the oxidized state, (iii) an electron transfer agent, (iv) a (D)-2-hydroxyglutarate dehydrogenase enzyme, and (v) a cofactor; and b) detecting (D)-2-hydroxyglutarate by measuring the production of the reduced state of the dye, wherein the presence of (D)-2-hydroxyglutarate in the sample of the subject is indicative for a mutation in an isocitrate dehydrogenase (IDH) gene or in a (D)-2-hydroxy-glutarate dehydrogenase gene in said subject. - View Dependent Claims (6)
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15. A kit comprising (i) a solvent, (ii) a dye having an oxidized state and a reduced state, wherein the reduced state can be distinguished from the oxidized state and wherein the dye is initially present in the oxidized state, (iii) an electron transfer agent, a (D)-2-hydroxy-glutarate dehydrogenase enzyme, and a cofactor.
Specification