title: Multiplex newborn screening for Pompe, Fabry, Hunter, Gaucher, and Hurler diseases using a digital microfluidic platform.
abstract: OBJECTIVE
  New therapies for lysosomal storage diseases (LSDs) have generated interest in screening newborns for these conditions. We present performance validation data on a digital microfluidic platform that performs multiplex enzymatic assays for Pompe, Fabry, Hunter, Gaucher, and Hurler diseases.
  
  
  METHODS
  We developed an investigational disposable digital microfluidic cartridge that uses a single dried blood spot (DBS) punch for performing a 5-plex fluorometric enzymatic assay on up to 44 DBS samples. Precision and linearity of the assays were determined by analyzing quality control DBS samples; clinical performance was determined by analyzing 600 presumed normal and known affected samples (12 for Pompe, 7 for Fabry and 10 each for Hunter, Gaucher and Hurler).
  
  
  RESULTS
  Overall coefficient of variation (CV) values between cartridges, days, instruments, and operators ranged from 2 to 21%; linearity correlation coefficients were ≥0.98 for all assays. The multiplex enzymatic assay performed from a single DBS punch was able to discriminate presumed normal from known affected samples for 5 LSDs.
  
  
  CONCLUSIONS
  Digital microfluidic technology shows potential for rapid, high-throughput screening for 5 LSDs in a newborn screening laboratory environment. Sample preparation to enzymatic activity on each cartridge is less than 3h.
authors: Advanced Liquid Logic, Inc., P.O. Box 14025, Research Triangle Park, NC 27709, USA.
authors: Ramakrishna S
authors: RS
authors: Sista
authors: Tong
authors: T
authors: Wang
authors: Ning
authors: N
authors: Wu
authors: Carrie
authors: C
authors: Graham
authors: Allen
authors: A
authors: Eckhardt
authors: Theodore
authors: T
authors: Winger
authors: Vijay
authors: V
authors: Srinivasan
authors: Deeksha
authors: D
authors: Bali
authors: David S
authors: DS
authors: Millington
authors: Vamsee K
authors: VK
authors: Pamula
mesh_terms: D059788:Dried Blood Spot Testing; D057075:Enzyme Assays; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D006801:Humans; D007231:Infant, Newborn; D016464:Lysosomal Storage Diseases / Q000097:blood / Q000145:classification / Q000175:diagnosis*; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D015997:Neonatal Screening; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D000520:alpha-Glucosidases / Q000097:blood* / Q000172:deficiency
chemical_list: D000520:alpha-Glucosidases
keywords: 4-MU; 4-MU-α-Gal; 4-MU-α-IDS; 4-MU-α-IDU; 4-MU-α-gluc; 4-MU-β-Gluc; 4-methyl umbelliferone; 4-methylumbelliferyl α-d-galactopyranoside; 4-methylumbelliferyl α-d-glucopyranoside; 4-methylumbelliferyl α-l-iduronate-2-sulfate; 4-methylumbelliferyl α-l-iduronide; 4-methylumbelliferyl β-d-glucopyranoside; ALL; CDC; CLSI; Centers for Disease Control and Prevention; Clinical and Laboratory Standards Institute; DBS; DMSO; Digital microfluidics; Dried blood spot; GAA; GBA; GLA; GalNac; High throughput; IDS; IDU; LSDs; Lysosomal storage disease; Multiplex enzymatic assay; N-acetyl-d-galactosamine; NBS; Newborn screening; QCBP; QCH; QCL; QCM; RFU; acid α-galactosidase; acid α-glucosidase; acid α-l-iduronate-2-sulfatase; acid α-l-iduronidase; acid β-d-glucosidase; advanced liquid logic, Inc.; d-Sac; d-saccharic acid 1,4 lactone; dimethyl sulfoxide; dried blood spot; lysosomal storage diseases; methyl-β-cyclodextrin; newborn screening; quality control base pool, 0% cord blood+100% leukoreduced adult blood adjusted to 50.5% hematocrit on filter paper; quality control high, cord blood adjusted to 50.5% hematocrit on filter paper; quality control low, 5% cord blood+95% leukoreduced adult blood adjusted to 50.5% hematocrit on filter paper; quality control medium, 50% cord blood+50% leukoreduced adult blood adjusted to 50.5% hematocrit on filter paper; relative fluorescence units; β-MBCD
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