Digital Microfluidic Platform for Multiplexing Enzyme Assays: Implications for Lysosomal Storage Disease Screening in Newborns
| Autor: | Scott Norton, Tong Wang, Carrie Graham, Vijay Srinivasan, Adviye A. Tolun, Deeksha Bali, Vamsee K. Pamula, Ramakrishna Sista, David S. Millington, Allen E. Eckhardt, Jeremy Rouse, Michael G. Pollack |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Clinical Biochemistry
Microfluidics Reference laboratory Article Neonatal Screening Lab-On-A-Chip Devices Lysosomal storage disease medicine Humans Fluorometry Digital microfluidics Dried blood Newborn screening Chromatography biology Chemistry Glycogen Storage Disease Type II Biochemistry (medical) Infant Newborn alpha-Glucosidases Clinical Enzyme Tests Microfluidic Analytical Techniques medicine.disease Molecular biology Substrate concentration Enzyme assay alpha-Galactosidase biology.protein Fabry Disease |
| Zdroj: | Clin Chem |
| Popis: | BACKGROUND Newborn screening for lysosomal storage diseases (LSDs) has been gaining considerable interest owing to the availability of enzyme replacement therapies. We present a digital microfluidic platform to perform rapid, multiplexed enzymatic analysis of acid α-glucosidase (GAA) and acid α-galactosidase to screen for Pompe and Fabry disorders. The results were compared with those obtained using standard fluorometric methods. METHODS We performed bench-based, fluorometric enzymatic analysis on 60 deidentified newborn dried blood spots (DBSs), plus 10 Pompe-affected and 11 Fabry-affected samples, at Duke Biochemical Genetics Laboratory using a 3-mm punch for each assay and an incubation time of 20 h. We used a digital microfluidic platform to automate fluorometric enzymatic assays at Advanced Liquid Logic Inc. using extract from a single punch for both assays, with an incubation time of 6 h. Assays were also performed with an incubation time of 1 h. RESULTS Assay results were generally comparable, although mean enzymatic activity for GAA using microfluidics was approximately 3 times higher than that obtained using bench-based methods, which could be attributed to higher substrate concentration. Clear separation was observed between the normal and affected samples at both 6- and 1-h incubation times using digital microfluidics. CONCLUSIONS A digital microfluidic platform compared favorably with a clinical reference laboratory to perform enzymatic analysis in DBSs for Pompe and Fabry disorders. This platform presents a new technology for a newborn screening laboratory to screen LSDs by fully automating all the liquid-handling operations in an inexpensive system, providing rapid results. |
| Databáze: | OpenAIRE |
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