Multiplex newborn screening for Pompe, Fabry, Hunter, Gaucher, and Hurler diseases using a digital microfluidic platform.
Multiplex newborn screening for Pompe, Fabry, Hunter, Gaucher, and Hurler diseases using a digital microfluidic platform.
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DOI：
10.1016/j.cca.2013.05.001
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发表时间：
2013-09-23
期刊：
影响因子：
--
通讯作者：
Pamula VK
中科院分区：
文献类型：
--
作者：
Sista RS;Wang T;Wu N;Graham C;Eckhardt A;Winger T;Srinivasan V;Bali D;Millington DS;Pamula VK
溶酶体贮积病 (LSD) 的新疗法引起了人们对新生儿筛查这些疾病的兴趣。我们在数字微流体平台上提供了性能验证数据，该平台对庞贝病、法布里病、亨特病、戈谢病和 Hurler 病进行多重酶测定。我们开发了一种研究性一次性数字微流体盒，它使用单个干血点 (DBS) 打孔器对多达 44 个 DBS 样品进行 5 重荧光酶测定。通过分析质量控制 DBS 样品来确定测定的精度和线性度；通过分析 600 个假定正常和已知受影响的样本（Pompe 12 个，Fabry 7 个，Hunter、Gaucher 和 Hurler 各 10 个）来确定临床表现。墨盒、天数、仪器和操作员之间的总体变异系数 (CV) 值在 2% 到 21% 之间；所有测定的线性相关系数均≥ 0.98。从单个 DBS 打孔机进行的多重酶测定能够区分 5 个 LSD 的假定正常样本和已知受影响样本。数字微流体技术显示出在新生儿筛查实验室环境中快速、高通量筛查 5 种 LSD 的潜力。每个柱上从样品制备到酶活性的时间不到 3 小时。
New therapies for lysosomal storage diseases (LSDs) have generated interest in screening newborns for these conditions. We present performance validation data on a digital microfluidic platform that performs multiplex enzymatic assays for Pompe, Fabry, Hunter, Gaucher, and Hurler diseases. We developed an investigational disposable digital microfluidic cartridge that uses a single dried blood spot (DBS) punch for performing a 5-plex fluorometric enzymatic assay on up to 44 DBS samples. Precision and linearity of the assays were determined by analyzing quality control DBS samples; clinical performance was determined by analyzing 600 presumed normal and known affected samples (12 for Pompe, 7 for Fabry and 10 each for Hunter, Gaucher and Hurler). Overall coefficient of variation (CV) values between cartridges, days, instruments, and operators ranged from 2 to 21%; linearity correlation coefficients were ≥ 0.98 for all assays. The multiplex enzymatic assay performed from a single DBS punch was able to discriminate presumed normal from known affected samples for 5 LSDs. Digital microfluidic technology shows potential for rapid, high-throughput screening for 5 LSDs in a newborn screening laboratory environment. Sample preparation to enzymatic activity on each cartridge is less than 3 hours.
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影响因子：
3.4
作者：
Millington DS;Sista R;Eckhardt A;Rouse J;Bali D;Goldberg R;Cotten M;Buckley R;Pamula V
通讯作者：
Pamula V
影响因子：
9.3
作者：
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影响因子：
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影响因子：
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Mechtler, Thomas P.;Stary, Susanne;Kasper, David C.
通讯作者：
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影响因子：
9.3
作者：
Sista, Ramakrishna S.;Eckhardt, Allen E.;Pamula, Vamsee K.
通讯作者：
Pamula, Vamsee K.