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CN104145022A 测定(D)-2-羟基戊二酸(D2HG)或(D)-2-羟基己二酸的工具和方法 有权

测定(D)-2-羟基戊二酸(D2HG)或(D)-2-羟基己二酸的工具和方法
Abstract:
本发明涉及检测样品中的(D)-2-羟基戊二酸或(D)-2-羟基己二酸的方法，所述方法包括步骤为：a)将样品与试剂混合物接触，其中所述试剂混合物包含:(i)溶剂,(ii)具有氧化态和还原态的染料,其中还原态可以与氧化态区分并且其中染料初始以氧化态存在，(iii)电子转移剂，(iv)(D)-2-羟基戊二酸脱氢酶，和(v)辅因子；和b)通过测量染料的还原态的生产检测(D)-2-羟基戊二酸或(D)-2-羟基己二酸。本发明还涉及用于诊断和/或监控对象中(D)-2-羟基戊二酸相关的疾病的方法。本发明也涵盖用于诊断对象中的异柠檬酸脱氢酶(IDH)基因中或(D)-2-羟基戊二酸(D2HG)脱氢酶基因中的突变的方法。此外，本发明提供了试剂盒，所述试剂盒包含(i)溶剂,(ii)具有氧化态和还原态的染料,其中还原态可以与氧化态区分并且其中染料初始以氧化态存在，(iii)电子转移剂，(iv)(D)-2-羟基戊二酸脱氢酶，和(v)辅因子。
Abstract(English):

The present invention relates to a method for detecting (D)-2-hydroxyglutarate 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 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-hydroxyglutarate-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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