Lavoie Pamela, Boutin Michel, Abaoui Mona, Auray-Blais Christiane
Division of Medical Genetics, Department of Pediatrics, Faculty of Medicine and Health Sciences, Université de Sherbrooke, Sherbrooke, Quebec, Canada.
Curr Protoc Hum Genet. 2016 Jul 1;90:17.22.1-17.22.12. doi: 10.1002/cphg.1.
Fabry disease is an X-linked lysosomal storage disorder caused by the absence or reduction of the enzyme α-galactosidase A activity. Currently, globotriaosylsphingosine (lyso-Gb3 ) and globotriaosylceramide (Gb3 ) are used as biomarkers to diagnose and monitor Fabry patients. However, recent metabolomic studies have shown that several glycosphingolipids are also elevated in biological fluids of affected patients and may be related to disease manifestations. This unit describes a multiplex methodology targeting the analysis of urinary lyso-Gb3 and seven structurally related analogs. A solid-phase extraction process is performed, then lyso-Gb3 and its analogs are analyzed simultaneously with an internal standard by ultra-performance liquid chromatography (UPLC) coupled to a tandem mass spectrometry (MS/MS) system. This methodology can be useful for the diagnosis of Fabry patients, including patients with cardiac variant mutations, but also to monitor the efficacy of therapeutic interventions, considering that lyso-Gb3 analogs are more elevated than lyso-Gb3 itself in urine. © 2016 by John Wiley & Sons, Inc.
法布里病是一种X连锁溶酶体贮积症，由α-半乳糖苷酶A活性缺失或降低引起。目前，球三糖基鞘氨醇（溶血型Gb3）和球三糖基神经酰胺（Gb3）被用作诊断和监测法布里病患者的生物标志物。然而，最近的代谢组学研究表明，几种糖鞘脂在受影响患者的生物体液中也有所升高，并且可能与疾病表现有关。本单元描述了一种针对尿液中溶血型Gb3和七种结构相关类似物进行分析的多重方法。先进行固相萃取过程，然后通过超高效液相色谱（UPLC）与串联质谱（MS/MS）系统联用，将溶血型Gb3及其类似物与内标同时进行分析。考虑到尿液中溶血型Gb3类似物比溶血型Gb3本身升高得更多，这种方法对于法布里病患者（包括具有心脏变异突变的患者）的诊断以及监测治疗干预的效果可能会很有用。© 2016约翰威立国际出版公司
