Development of a new tandem mass spectrometry method for urine and amniotic fluid screening of oligosaccharidoses
Development of a new tandem mass spectrometry method for urine and amniotic fluid screening of oligosaccharidoses
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DOI：
10.1002/rcm.7860
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发表时间：
2017-06-15
影响因子：
2
通讯作者：
Froissart, Roseline
中科院分区：
文献类型：
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作者：
Piraud, Monique;Pettazzoni, Magali;Froissart, Roseline
理由：诊断低聚糖病的第一步是检测尿液中是否有异常低聚糖，通常采用薄层色谱（TLC）法进行检测，薄层色谱法灵敏度低，但效率高。开发一种串联质谱（MS/MS）技术有望取代薄层色谱。方法：最近使用基质辅助激光解吸/电离飞行时间质谱法研究了不正常的低聚糖，从而可以明确地鉴定低聚糖。在此基础上，我们建立了一种高效的液相色谱(LC)/MS/MS分析方法，使用更常见的三重四极杆串联质谱仪进行低聚糖分析。结果：对低聚糖（n = 97）和对照组（n = 240）尿样进行了分析。用这种方法得到了每种寡糖病的特定模式。在尿液中，它不仅可以识别以前通过TLC识别的所有低聚糖病（focusidosis, alphamannosidosis, aspartylglucosaminuria， GM1神经节脂病，唾液病，半乳糖唾液病和Schindler病），而且还可以将诊断领域扩展到II型粘脂病，Sandhoff病和β -甘露糖病。同样的技术应用于16例寡糖病胎儿的羊水上清（n = 16），与37例未受影响的胎儿进行比较。所有受影响的胎儿均可明确识别：唾液中毒（n = 3）、半乳糖唾液中毒（n = 4）、天冬氨酸糖氨基尿症（n = 1）、II型粘脂中毒（n = 4）或GM1神经节脂中毒（n = 4）。该技术可用于产前早期诊断，也可用于非免疫性积水胎儿的寡糖病筛查。结论：该方法快速、简便，每个样品的LC分析时间为13 min。在没有特定标准品和每种化合物的标记内标品的情况下，无法进行定量验证。即使这种LC/MS/MS方法只是定性的，但它的特异性很强，比TLC灵敏得多。它允许低聚糖的尿筛选，甚至轻度或迟发形式，并筛选产前形式的羊水。版权所有：John Wiley & Sons， Ltd。
RATIONALE: The first step in the diagnosis of oligosaccharidoses is to evidence abnormal oligosaccharides excreted in urine, usually performed by the poorly sensitive but efficient thin layer chromatography (TLC) method. Developing a tandem mass spectrometry (MS/MS) technique could be of great interest to replace TLC.METHODS: Abnormal underivatized oligosaccharides have been recently studied using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, allowing the unambiguous identification of oligosaccharidoses. Based on this previous work, we developed an advantageous and efficient liquid chromatography (LC)/MS/MS method using a more common triple quadrupole tandem mass spectrometer for oligosaccharides analysis.RESULTS: Oligosaccharidoses (n = 97) and control (n = 240) urine samples were analysed. A specific pattern was obtained for each oligosaccharidosis using this method. In urine, it allows not only the identification of all the oligosaccharidoses previously identified by TLC (fucosidosis, alphamannosidosis, aspartylglucosaminuria, GM1 gangliosidosis, sialidosis, galactosialidosis and Schindler disease), but also extends the field of diagnosis to mucolipidosis type II, Sandhoff disease, and beta-mannosidosis. The same technique was applied to 16 amniotic fluid supernatants from oligosaccharidosis-affected foetuses (n = 16) compared with 37 unaffected. All the affected foetuses could be clearly identified: sialidosis (n = 3), galactosialidosis (n = 4), aspartylglucosaminuria (n = 1), mucolipidosis type II (n = 4) or GM1 gangliosidosis (n = 4). This technique can be applied to early prenatal diagnosis as well as to the oligosaccharidosis screening in the case of non-immune hydrops fetalis.CONCLUSIONS: The method is quick and easy to run, with an LC analysis time of 13 min per sample. The quantitative validation could not be obtained in the absence of a specific standard and of a labelled internal standard for each compound. Even if this LC/MS/MS method is only qualitative, it is very specific and much more sensitive than TLC. It allows the urinary screening of oligosaccharidoses, even mild or late-onset forms, and the screening of antenatal forms in amniotic fluid. Copyright (C) 2017 John Wiley & Sons, Ltd.