Digital microfluidic platform for multiplexing enzyme assays: implications for lysosomal storage disease screening in newborns.

Journal Article (Journal Article)

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.

Full Text

Duke Authors

Cited Authors

  • Sista, RS; Eckhardt, AE; Wang, T; Graham, C; Rouse, JL; Norton, SM; Srinivasan, V; Pollack, MG; Tolun, AA; Bali, D; Millington, DS; Pamula, VK

Published Date

  • October 2011

Published In

Volume / Issue

  • 57 / 10

Start / End Page

  • 1444 - 1451

PubMed ID

  • 21859904

Pubmed Central ID

  • PMC8917906

Electronic International Standard Serial Number (EISSN)

  • 1530-8561

Digital Object Identifier (DOI)

  • 10.1373/clinchem.2011.163139


  • eng

Conference Location

  • England