Timely diagnosis is essential for patients with neurometabolic disorders to enable targeted treatment. Next‐Generation Metabolic Screening (NGMS) allows for simultaneous screening of multiple diseases and yields a holistic view of disturbed metabolic pathways. We applied this technique to define a cerebrospinal fluid (CSF) reference metabolome and validated our approach with patients with known neurometabolic disorders. Samples were measured using ultra‐high‐performance liquid chromatography‐quadrupole time‐of‐flight mass spectrometry followed by (un)targeted analysis. For the reference metabolome, CSF samples from patients with normal general chemistry results and no neurometabolic diagnosis were selected and grouped based on sex and age (0‐2/2‐5/5‐10/10‐15 years). We checked the levels of known biomarkers in CSF from seven patients with five different neurometabolic disorders to confirm the suitability of our method for diagnosis. Untargeted analysis of 87 control CSF samples yielded 8036 features for semiquantitative analysis. No sex differences were found, but 1782 features (22%) were different between age groups (q < 0.05). We identified 206 diagnostic metabolites in targeted analysis. In a subset of 20 high‐intensity metabolites and 10 biomarkers, 17 (57%) were age‐dependent. For each neurometabolic patient, ≥1 specific biomarker(s) could be identified in CSF, thus confirming the diagnosis. In two cases, age‐matching was essential for correct interpretation of the metabolomic profile. In conclusion, NGMS in CSF is a powerful tool in defining a diagnosis for neurometabolic disorders. Using our database with many (age‐dependent) features in CSF, our untargeted approach will facilitate biomarker discovery and further understanding of mechanisms of neurometabolic disorders.

中文翻译：

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使用年龄依赖性脑脊液代谢组学特征确认神经代谢诊断。

及时诊断对于神经代谢紊乱患者至关重要，以便进行针对性治疗。下一代代谢筛查 (NGMS) 允许同时筛查多种疾病，并产生紊乱代谢途径的整体视图。我们应用这种技术来定义脑脊液 (CSF) 参考代谢组，并在患有已知神经代谢疾病的患者中验证了我们的方法。使用超高效液相色谱-四极杆飞行时间质谱法测量样品，然后进行（非）靶向分析。对于参考代谢组，根据性别和年龄（0-2/2-5/5-10/10-15 岁）选择和分组来自一般化学结果正常且无神经代谢诊断的患者的 CSF 样本。我们检查了来自七名患有五种不同神经代谢疾病的患者的脑脊液中已知生物标志物的水平，以确认我们的诊断方法的适用性。87 个对照 CSF 样本的非靶向分析产生了 8036 个用于半定量分析的特征。没有发现性别差异，但 1782 个特征（22%）在年龄组之间存在差异（q  < 0.05）。我们在靶向分析中鉴定了 206 种诊断代谢物。在 20 种高强度代谢物和 10 种生物标志物的子集中，17 种（57%）具有年龄依赖性。对于每位神经代谢患者，可在 CSF 中鉴定出≥1 个特定生物标志物，从而确认诊断。在两种情况下，年龄匹配对于正确解释代谢组学特征至关重要。总之，脑脊液中的 NGMS 是确定神经代谢疾病诊断的有力工具。使用我们的数据库在 CSF 中具有许多（年龄相关）特征，我们的非靶向方法将促进生物标志物的发现和对神经代谢紊乱机制的进一步了解。