Multiplex Analysis of Inborn Errors of Metabolism

本项目的总体目标是开发用于新生儿的串联质谱 筛选存在治疗选择的溶酶体贮积病亚类 或正在开发中。新生儿筛查卡上的干血斑上的一个穿孔， 用作溶酶体酶的来源，并将其与一系列 在合适的缓冲液中的底物，以允许形成产物。第二冲头用于 用有机溶剂提取以回收溶酶体贮积病的生物标志物。两 测定可以组合成单个多重串联质谱测定 新生儿筛查多种可治疗的溶酶体贮积病。我们建议试点 这种高度多重检测在华盛顿州新生儿筛查实验室进行。我们 还将对干血斑进行外显子组DNA测序， 低于临界值的活动。这些数据将使我们能够确定 值和筛选测定的假阳性率。这些试点研究将探讨 新生儿筛查可治疗的溶酶体储积病的可行性。第二 该项目的组成部分是开发串联质谱分析，以支持后， 新生儿溶酶体贮积症筛查分析。我们将使用高分辨率 测定血液来源的白细胞中酶水平的微小变化， 可以估计溶酶体贮积病的严重程度。

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.