Multiplex Analysis of Inborn Errors of Metabolism

先天性代谢缺陷的多重分析

• 批准号: 9923620
• 负责人: Michael H Gelb
• 金额: $ 55.36万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9923620
• 关键词: Arylsulfatases; Bile Acids; Biological Assay; Biological Markers; Birth; Blood; Buffers; Cerebrotendinous Xanthomatosis; Clinical; Clinical Trials; Collection; DNA sequencing; Data; Development; Diagnosis; Disease; Enzymes; Fabry Disease; Genotype; Goals; Grant; Inborn Errors of Metabolism; Incubated; Isotope Labeling; Laboratories; Leukocytes; Life; Lysosomal Storage Diseases; Measures; Metachromatic Leukodystrophy; Methods; Mucopolysaccharidoses; Mucopolysaccharidosis I; Neonatal Screening; Neuronal Ceroid-Lipofuscinosis; Newborn Infant; Organic solvent product; Phase; Pilot Projects; Predictive Value; Research; Resolution; Sampling; Severities; Source; Spottings; Technology; Testing; Treatment outcome; Washington; base; enzyme activity; enzyme deficiency; exome; improved; interest; screening; tandem mass spectrometry; therapy development; thioesterase PPT1 gene product

The overall goal of this project is develop tandem mass spectrometry for newborn screening of the subset of lysosomal storage diseases for which treatment options exist or are being developed. A punch from a dried blood spot on a newborn screening card is used as a source of lysosomal enzymes, and this is incubated with a collection of substrates in a suitable buffer to allow formation of products. A second punch is used for extraction with organic solvent to recover biomarkers for lysosomal storage diseases. Both assays can be combined into a single multiplex tandem mass spectrometry assay for newborn screening of multiple treatable lysosomal storage diseases. We propose to pilot this highly multiplexed assay in the Washington State Newborn Screening Laboratory. We will also carry out exome DNA sequencing on dried blood spots that give an enzyme activity below the cut-off value. This data will allow us to determine the positive predictive values and false positive rates for the screening assay. These pilot studies will explore the feasibility of newborn screening of treatable lysosomal storage diseases. The second component of the project is to develop tandem mass spectrometry assays to support post- newborn screening analysis for lysosomal storage diseases. We will use high resolution assays to measure small changes in enzyme levels in blood-derived leukocytes so that the severity of the lysosomal storage disease can be estimated.

该项目的总体目标是开发用于新生儿的串联质谱学 筛选存在治疗选择的溶酶体储存性疾病的子集 或正在开发中。从新生儿筛查卡上的干血点打出的一拳是 用作溶酶体酶的来源，这是与一系列 在合适的缓冲液中放置衬底，以允许形成产品。第二个冲头用于 有机溶剂萃取法回收溶酶体储存疾病的生物标志物。两者都有 分析可以合并到单个多路串联质谱分析中 新生儿多种可治疗溶酶体贮积性疾病的筛查。我们建议试行 华盛顿州新生儿筛查实验室的这项高度多元化的检测。我们 还将对干燥的血斑进行外显子DNA测序，这些斑点提供了一种酶 低于分界值的活动。这些数据将使我们能够确定积极的预测 筛查试验的数值和假阳性率。这些初步研究将探索 新生儿可治疗溶酶体储存疾病筛查的可行性。第二 该项目的组成部分是开发串联质谱分析，以支持后 新生儿溶酶体贮积性疾病筛查分析。我们将使用高分辨率 测量血源性白细胞中酶水平的微小变化的检测方法 溶酶体储存病的严重程度是可以估计的。