Digital Microfluidic Platform for Multiplexing Enzyme Assays: Implications for Lysosomal Storage Disease Screening in Newborns

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 t...

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Bibliographic Details
Published in:Clinical chemistry (Baltimore, Md.) Vol. 57; no. 10; pp. 1444 - 1451
Main Authors: Sista, Ramakrishna S, Eckhardt, Allen E, Wang, Tong, Graham, Carrie, Rouse, Jeremy L, Norton, Scott M, Srinivasan, Vijay, Pollack, Michael G, Tolun, Adviye A, Bali, Deeksha, Millington, David S, Pamula, Vamsee K
Format: Journal Article
Language:English
Published: Washington, DC American Association for Clinical Chemistry 01.10.2011
Oxford University Press
Subjects:
alpha-Galactosidase - blood
alpha-Glucosidases - blood
Analytical, structural and metabolic biochemistry
Automation
Biological and medical sciences
Clinical Enzyme Tests - instrumentation
Disease
Enzymatic activity
Fabry Disease - diagnosis
Fluorometry
Fundamental and applied biological sciences. Psychology
Genetics
Glycogen Storage Disease Type II - diagnosis
Humans
Infant, Newborn
Investigative techniques, diagnostic techniques (general aspects)
Lab-On-A-Chip Devices
Medical sciences
Medical screening
Methods
Microfluidic Analytical Techniques - instrumentation
Molecular biophysics
Neonatal Screening
Public health
Human
Assay
Multiplexing
Newborn
Enzyme
Clinical biology
Biochemistry
Molecular biology
Medical screening
Lysosomal storage disease
ISSN:0009-9147, 1530-8561, 1530-8561
Online Access:Get full text
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Summary: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. 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. 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. 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.
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Author Contributions: All authors confirmed they have contributed to the intellectual content of this paper and have met the following 3 requirements: (a) significant contributions to the conception and design, acquisition of data, or analysis and interpretation of data; (b) drafting or revising the article for intellectual content; and (c) final approval of the published article.
ISSN:0009-9147
1530-8561
1530-8561
DOI:10.1373/clinchem.2011.163139