Short-incubation mass spectrometry assay for lysosomal storage disorders in newborn and high-risk population screening
Short-incubation mass spectrometry assay for lysosomal storage disorders in newborn and high-risk population screening
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DOI：
10.1016/j.jchromb.2012.09.012
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发表时间：
2012-11-01
影响因子：
3
通讯作者：
Kasper, David C.
中科院分区：
文献类型：
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作者：
Mechtler, Thomas P.;Metz, Thomas F.;Kasper, David C.
在过去的几年中，由于新的治疗策略的可用性加上诊断技术的发展，对新生儿和高危人群中溶酶体贮积症（LSD）的早期检测策略的兴趣有所增加。我们报告了一种基于短孵育质谱的方案的开发，该方案允许在4小时内检测戈谢病、尼曼-匹克A/B病、庞贝氏病、法布里病和粘多糖样变性I型疾病，包括从干血斑制备样品。优化的样品处理无需耗时的离线制备，如液-液和固相萃取，允许使用底物盒和氘代内标物同时定量五种溶酶体酶的活性。应用3小时的孵育时间显示，所有五种酶活性的日内CV%值范围分别为4%至11%。在第一次临床评价中，我们测试了825名未受影响的新生儿和16例LSD患者使用多路复用，湍流色谱-超高效液相色谱-串联质谱仪测定。所有受影响的患者都得到了准确的识别，并且可以与未受影响的新生儿区分开来。与先前发表的包括过夜孵育的两天测定相比，该方案能够在半天内检测从样品到第一个结果的溶酶体酶活性。(C)2012 Elsevier BM. All rights reserved.
The interest in early detection strategies for lysosomal storage disorders (LSDs) in newborns and high-risk population has increased in the last years due to the availability of novel treatment strategies coupled with the development of diagnostic techniques. We report the development of a short-incubation mass spectrometry-based protocol that allows the detection of Gaucher, Niemann-Pick A/B, Pompe, Fabry and mucopolysaccharidosis type I disease within 4h including sample preparation from dried blood spots. Optimized sample handling without the need of time-consuming offline preparations, such as liquid-liquid and solid-phase extraction, allows the simultaneous quantification of five lysosomal enzyme activities using a cassette of substrates and deuterated internal standards. Applying incubation times of 3 h revealed in intra-day CV% values ranging from 4% to 11% for all five enzyme activities, respectively. In a first clinical evaluation, we tested 825 unaffected newborns and 16 patients with LSDs using a multiplexed, turbulent flow chromatography-ultra high performance liquid chromatography-tandem mass spectrometer assay. All affected patients were identified accurately and could be differentiated from non-affected newborns. In comparison to previously published two-day assays, which included an overnight incubation, this protocol enabled the detection of lysosomal enzyme activities from sample to first result within half a day. (C) 2012 Elsevier BM. All rights reserved.