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
Language
- eng
Conference Location
- England