Multiplex Analysis of Inborn Errors of Metabolism

先天性代谢缺陷的多重分析

• 批准号: 7143170
• 负责人: FRANTISEK TURECEK
• 金额: $ 30.23万
• 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 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7143170
• 关键词: acyl group; aminoacid; cell line; clinical research; diagnosis design /evaluation; electrospray ionization mass spectrometry; enzyme activity; heme; human subject; inborn lysosomal enzyme disorder; inborn metabolism disorder; metachromatic leukodystrophy; method development; mucopolysaccharidosis; organic acid; patient oriented research; pediatrics; porphyrias; protein biosynthesis

DESCRIPTION (provided by applicant): The unifying goal of our proposed studies is to further develop tandem mass spectrometry (MS) as an analytical method for the multiplex analysis of enzyme activities of diagnostic value for the detection of inborn errors of metabolism. Tandem MS assays will be developed for the enzymes relevant to the lysosomal storage diseases (LSD) belonging to the Mucopolysaccharidosis groups II (MPS-II, Hunter), MPSVI (Maroteaux-Lamy), and metachromatic leukodystrophy, using dried blood spots on newborn screening cards as the enzyme source. Treatment of these disorders is in late-stage development, and our assays will make it possible for newborn screening laboratories to spot these diseases prior to the development of irreversible phenotypic abnormalities. Another group of diseases to be tackled is the Sanfilippo syndromes A-D (Mucopolysaccharidosis IIIA-D). The biochemical analysis of Sanfilippo syndromes is difficult because the same phenotypic symptoms present in patients when one of the four different enzymes is deficient. Furthermore, previous assays of the relevant enzymes have been difficult owing to the need to use a collection of different assay techniques. Tandem MS assays of all four enzymes relevant to Sanfilippo syndrome will use the same single analytical platform as for the other LSD. We will develop assays for enzymes in the heme biosynthetic pathway for the biochemical diagnosis of the various forms of porphyrias. Porphyrias are not typically assayed in most clinical laboratories because of the need for highly specialized protocols. We will attempt to develop tandem MS assays for all of the individual enzymes in the heme biosynthetic pathway so that the assay can be carried out in a larger number of laboratories, which will help physicians diagnose this set of disorders.

描述（由申请人提供）： 我们提出的研究的统一目标是进一步开发串联质谱（MS）作为一种分析方法，用于对具有诊断价值的酶活性进行多重分析，以检测先天性代谢错误。将针对属于粘多糖贮积症 II 组（MPS-II，Hunter）、MPSVI（Maroteaux-Lamy）和异染性脑白质营养不良的溶酶体贮积病 (LSD) 相关的酶开发串联 MS 检测，使用新生儿筛查中的干血斑卡作为酶源。这些疾病的治疗处于后期开发阶段，我们的检测将使新生儿筛查实验室能够在出现不可逆表型异常之前发现这些疾病。另一组需要解决的疾病是 Sanfilippo 综合征 A-D（粘多糖贮积症 IIIA-D）。 Sanfilippo 综合征的生化分析很困难，因为当四种不同酶中的一种缺乏时，患者会出现相同的表型症状。此外，由于需要使用不同测定技术的集合，以前对相关酶的测定一直很困难。与 Sanfilippo 综合征相关的所有四种酶的串联 MS 检测将使用与其他 LSD 相同的单一分析平台。我们将开发血红素生物合成途径中酶的检测方法，以对各种形式的卟啉症进行生化诊断。由于需要高度专业化的方案，大多数临床实验室通常不会对卟啉症进行检测。我们将尝试开发针对血红素生物合成途径中所有单个酶的串联质谱检测，以便该检测可以在更多的实验室中进行，这将有助于医生诊断这组疾病。