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...

Celý popis

Uloženo v:

Podrobná bibliografie
Vydáno v:	Clinical chemistry (Baltimore, Md.) Ročník 57; číslo 10; s. 1444 - 1451
Hlavní autoři:	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
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	Washington, DC American Association for Clinical Chemistry 01.10.2011 Oxford University Press
Témata:	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
On-line přístup:	Získat plný text
Tagy:	Přidat tag Žádné tagy, Buďte první, kdo vytvoří štítek k tomuto záznamu!

Popis
Shrnutí:	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.
Bibliografie:	ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 14 content type line 23 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