Gaucher and Niemann-Pick diseases - enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards
Gaucher and Niemann-Pick diseases - enzymatic diagnosis in dried blood spots on filter paper: retrospective diagnoses in newborn-screening cards
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DOI：
10.1016/s0009-8981(01)00798-7
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发表时间：
2002-03-01
影响因子：
5
通讯作者：
Casentini, C
中科院分区：
文献类型：
--
作者：
Chamoles, NA;Blanco, M;Casentini, C
背景资料：戈谢病（GD）和尼曼-皮克病（NP）分别由溶酶体酶酸性β-D-葡糖苷酶（ABG）和酸性鞘磷脂酶（ASM）的活性缺陷引起。为了诊断，通常在白细胞或培养的成纤维细胞的提取物中测量这些酶。壳三糖苷酶（CTE），一种几丁质分解酶，在戈谢病患者的血浆中显著增加。我们描述了用于测定酸性β-D-葡萄糖苷酶、酸性鞘磷脂酶、壳三糖苷酶和α-N-乙酰氨基半乳糖苷酶（NAGA）的方法，所述酶作为在滤纸上干燥的血斑中的对照酶。方法：在含有直径为3mm血斑的试管中加入洗脱液和底物溶液。在37 ℃下孵育后，将水解产物的量与校准品进行比较，以允许酶活性的定量。我们检查了80名健康对照、54名戈谢病患者、8名尼曼-匹克病患者、27名专性携带者以及来自一名戈谢病病例和一名尼曼-匹克病病例的新生儿筛查卡（NSC）。结果与结论：所描述的方法可用于使用足够稳定以通过邮件运输到检测实验室的样品来鉴定戈谢和尼曼-匹克患者和对照。两种疾病的诊断都明确建立在新的筛查卡上。生物材料中增加了新的免疫筛查卡，可以识别戈谢病和尼曼-匹克病患者。(C)2002 Elsevier Science B.V.保留所有权利。
Background: Gaucher disease (GD) and Niemann-Pick (NP) disease are caused by deficient activity of the lysosomal enzymes acid beta-D-glucosidase (ABG) and acid sphingomyelinase (ASM), respectively, For diagnosis, these enzymes are usually measured in the extracts of leukocytes or cultured fibroblasts. Chitotriosidase (CTE), a chitinolytic enzyme, is markedly increased in the plasma of Gaucher patients. We describe methods for the assay of acid beta-D-glucosidase, acid sphingomyelinase, chitotriosidase, and alpha-N-acetyl-galactosaminidase (NAGA) as a control enzyme in blood spots that were dried onto filter paper. Methods: To tubes containing a 3 mm-diameter blood spot, we added elution liquid and substrate solution. After incubation at 37 degreesC, the amount of hydrolyzed product was compared with a calibrator to allow the quantification of enzyme activity, We examined 80 healthy controls, 54 Gaucher patients, 8 Niemann-Pick patients, 27 obligate carriers, and the newborn-screening cards (NSC) from a case of Gaucher and a case of Niemann-Pick disease. Results and conclusion: The described methodology is useful to identify Gaucher and Niemann-Pick patients and controls, using samples that are sufficiently stable to be transported to the testing laboratory by mail. The diagnosis of both diseases on a newborn-screening card was clearly established. The newborn-screening card has been added to the biological materials that allow the identification of patients with Gaucher and Niemann-Pick diseases. (C) 2002 Elsevier Science B.V. All rights reserved.