A myeloperoxidase-specific assay based upon bromide-dependent chemiluminescence of luminol
A myeloperoxidase-specific assay based upon bromide-dependent chemiluminescence of luminol
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DOI：
10.1006/abio.1999.4206
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发表时间：
1999-08-15
影响因子：
2.9
通讯作者：
Birnboim, HC
中科院分区：
文献类型：
--
作者：
Haqqani, AS;Sandhu, JK;Birnboim, HC
髓过氧化物酶(MPO；EC 1.11.1.7)活性的测定常被用来作为中性粒细胞渗入组织的标志。然而，大多数MPO的酶分析容易受到组织中存在的其他过氧化物酶(包括嗜酸性粒细胞过氧化物酶，EPX)和血红蛋白(如血红蛋白和肌红蛋白)的干扰。在这份报告中，我们描述了一种使用鲁米诺作为化学发光探针的溴化物依赖化学发光(BR-CL)分析方法。该方法可以区分MPO反应和非特异性的过氧化物酶反应。MPO专一性反应分两步进行：(1)在pH为5的条件下，KBr2和H_2O_2酶促生成次溴酸(HOBR)；(2)HOBr和H_2O_2与鲁米诺发生自发反应，生成BR-CL信号。该检测方法具有足够的灵敏度，可以检测到MPO。
Measurement of myeloperoxidase (MPO; EC 1.11.1.7) activity is often used as a marker of neutrophil infiltration into tissues. However, most enzymatic assays for MPO are susceptible to interference from other peroxidases (including eosinophil peroxidase, EPX) and hemoproteins (such as hemoglobin and myoglobin) present in the tissues. In this report, we describe a bromide-dependent chemiluminescence (Br-CL) assay that uses luminol as a chemiluminescence probe. The assay can distinguish between MPO and nonspecific peroxidase reactions. The MPO-specific reaction is believed to proceed in two steps: (i) the enzymatic generation of hypobromous acid (HOBr) from KBr and H2O2 at pH 5 and (ii) the spontaneous reaction of HOBr and H2O2 with luminol to give a Br-CL signal. The assay is sufficiently sensitive to allow detection of MPO in