Departments of Chemistry, University of Washington, Seattle, WA 98195, USA.
Departments of Pediatrics, University of Washington, Seattle, WA 98195, USA.
Mol Genet Metab. 2018 Jun;124(2):101-108. doi: 10.1016/j.ymgme.2018.03.012. Epub 2018 Mar 29.
All States screen for biotinidase deficiency and galactosemia, and X-linked adrenoleukodystrophy (X-ALD) has recently been added to the Recommended Uniform Screening Panel (RUSP).We sought to consolidate these tests by combining them into a single multiplex tandem mass spectrometry assay as well as to improve the current protocol for newborn screening of galactosemia.A 3 mm punch of a dried blood spot (DBS) was extracted with organic solvent for analysis of the C26:0-lysophosphatidylcholine biomarker for X-ALD.An additional punch was used to assay galactose-1-phosphate uridyltransferase (GALT) and biotinidase.All assays were combined for a single injection for analysis by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (2.3 min per sample).The GALT LC-MS/MS assay does not give a false positive for galactosemia if glucose-6-phosphate dehydrogenase is deficient.The multiplex assay shows acceptable reproducibility and provides for rapid analysis of X-ALD, biotinidase deficiency, and galactosemia.The throughput and ease of sample preparation are acceptable for newborn screening laboratories.We also show that the LC-MS/MS assay is expandable to include several other diseases including Pompe and Hurler diseases (enzymatic activities and biomarkers).Because of consolidation of assays, less manpower is needed compared to running individual assays on separate platforms.The flexibility of the LC-MS/MS platform allows each newborn screening laboratory to analyze the set of diseases offered in their panel.
所有州都筛查生物素酶缺乏症和半乳糖血症，并且 X 连锁肾上腺脑白质营养不良（X-ALD）最近已被添加到推荐的统一筛选面板（RUSP）中。我们试图通过将这些测试组合到一个单一的多重串联质谱测定法中，同时改进当前的半乳糖血症新生儿筛查方案，来整合这些测试。从干燥血斑（DBS）中提取 3mm 的打孔样本，用有机溶剂分析 X-ALD 的 C26:0-溶血磷脂酰胆碱生物标志物。再使用一个打孔样本来测定半乳糖-1-磷酸尿苷转移酶（GALT）和生物素酶。所有检测均组合在单个注射中，通过液相色谱-串联质谱（LC-MS/MS）进行分析（每个样本 2.3 分钟）。如果葡萄糖-6-磷酸脱氢酶缺乏，GALT LC-MS/MS 检测对半乳糖血症不会产生假阳性。该多重检测具有可接受的重现性，可快速分析 X-ALD、生物素酶缺乏症和半乳糖血症。该方法的通量和样本制备的简便性可满足新生儿筛查实验室的要求。我们还表明，LC-MS/MS 测定法可扩展到包括其他几种疾病，包括庞贝病和Hurler 病（酶活性和生物标志物）。由于检测的整合，与在单独平台上运行单个检测相比，所需的人力更少。LC-MS/MS 平台的灵活性允许每个新生儿筛查实验室分析其面板中提供的一组疾病。
