SBIR-STTR Award

Newborn screening is currently performed by collecting dried bloodspots from infants and then sending them to a lab for analysis. There is an increasing necessity to screen for a number of disease conditions for which therapies are becoming available. Lysosomal storage diseases alone number greater than 40. There is a need to minimize the blood collected for screening purposes while maximizing the number of conditions screened. Also, conventional laboratory analysis of blood samples in a birthing center is impractical. An automated, inexpensive, multi-analyte and multiplexed newborn screening analyzer will help reduce the blood analysis volume, reduce the time-to-result, and make the analyzer available at the point-of- delivery (birthing centers). Based on Advanced Liquid Logic's previous demonstrations of digital microfluidic manipulation of nanoliter-sized droplets of enzymatic reagents and sample for dispensing from on-chip reservoirs, high speed transport, mixing, splitting and dilution, and absorbance, fluorescence, and luminescence detection of the assays, a disposable lab-on-a-chip will be developed. Ultimately, a disposable lab-on-a-chip for newborn screening will be developed where one punch from the dried blood spot or fresh blood will be the input to the system and it outputs the screening results in a completely automated fashion. Phase I work will focus on fabricating a microfluidic platform upon which microliters of dried blood spot extracts, reference standards, and reagent droplets will be dispensed, transported, mixed, assayed, and disposed. Fluorescence assays for determining activities of enzymes involved in Pompe, Fabry, and Hurler diseases will be transitioned to droplet-based format. Demonstration of these enzymatic assays establishes the feasibility of a digital microfluidic chip for newborn screening.Advanced Liquid Logic, Inc. is developing a lab-on-a-chip that integrates clinical chemistry, immunoassays, DNA amplification, and DNA sequencing on its proprietary digital (droplet-based) microfluidic platform for clinical diagnostics. The proposed project aims at demonstrating the feasibility of performing newborn screening for lysosomal storage disorders including Pompe, Fabry, and Hurler diseases from a single dried blood spot obtained from the heelstick of a newborn using a microfluidic lab-on-a-chip

Newborn screening is currently performed by collecting dried bloodspots from infants and then sending them to a lab for analysis. Lysosomal storage diseases alone number greater than 40 and there is an increasing necessity to screen for a number of disease conditions for which therapies are becoming available. Tandem mass spectrometry is an excellent multiplex detection technology widely utilized in newborn screening, but when applied to enzyme assays it is very expensive, time consuming, labor intensive, and more importantly the multiplexing power of mass spectrometry is not leveraged because each assay has to be performed individually. There is a need for an inexpensive, rapid, automated, and scalable technology for performing newborn screening assays that a mass spectrometer is not well-suited to perform. A digital microfluidic system for performing enzymatic assays in newborn screening will enable walkaway automation and multiplex several assays very inexpensively. Based on Advanced Liquid Logic's successful demonstration of digital microfluidic manipulation of nanoliter-sized droplets of enzymatic reagents and sample for dispensing from on-chip reservoirs, high speed transport, mixing, splitting and dilution, and absorbance, fluorescence, and luminescence detection of the assays, a disposable lab-on-a- chip will be developed. In phase I, we have successfully demonstrated a multiplex fluorescence enzymatic assay on a digital microfluidic cartridge to setup screening for Pompe, Fabry, and Hurler disorders on dried blood spot samples. Phase II work will focus on increasing the throughput of the cartridge to screen for 6 lysosomal storage disorders for which therapies exist including Pompe, Fabry, Hurler, Hunter, Gaucher, and Maroteaux-Lamy, on 96 dried blood spot samples yielding a total of 576 enzymatic assays on a single cartridge. Also, a pilot screening study will be performed in collaboration with Duke University and North Carolina State Lab of Public Health, which would involve screening about 10,000 dried blood spots on the digital microfluidic platform.

Public Health Relevance:
Newborn screening is performed on every infant born in the US and there is a growing interest in increasing the number of conditions screened for. In this project, a digital microfluidic platform will be developed to screen for many treatable conditions simultaneously using much lesser volume of blood from an infant. It would be greatly useful in identifying treatable diseases earlier.

Thesaurus Terms:
0-6 Weeks Old; Address; Assay; Automation; Bioassay; Biochemistry; Biologic Assays; Biological Assay; Blood; Blood Volume; Budgets; Chemistry, Biological; Collaborations; Data; Detection; Development; Disease; Disorder; Economics; Enzymes; Fluorescence; Goals; Head; Hemoglobin; Hereditary Metabolic Disorder; Inborn Errors Of Metabolism; Infant; Infant, Newborn; Lab On A Chip; Letters; Liquid Substance; Logic; Lysosomal Enzyme Disorders; Lysosomal Storage Diseases; Mass Spectrum; Mass Spectrum Analysis; Measurement; Metabolism, Inborn Errors; Microfluidic; Microfluidics; Neonatal Screening; Newborn Infant; Newborn Infant Screening; Newborns; North Carolina; Pharmaceutical Agent; Pharmaceuticals; Pharmacologic Substance; Pharmacological Substance; Phase; Photometry/Spectrum Analysis, Mass; Play; Price; Process; Programs (Pt); Programs [publication Type]; Public Health; Reagent; Research; Reticuloendothelial System, Blood; Role; Sampling; Screening Procedure; Spectrometry, Mass; Spectroscopy, Mass; Spectrum Analyses, Mass; Spectrum Analysis, Mass; Speed; Speed (Motion); Spottings; System; System, Loinc Axis 4; Technology; Testing; Time; Universities; Validation; Work; Assay Development; Base; Cost; Design; Designing; Digital; Disease/Disorder; Fluid; Inborn Lysosomal Enzyme Disorder; Inborn Metabolism Disorder; Instrument; Interest; Liquid; Luminescence; Mass Spectrometer; Meetings; Micro-Total Analysis System; Mu-Tas; Multiplex Detection; Nano Litre; Nanoliter; Nanolitre; Newborn Human (0-6 Weeks); Newborn Screening; Pricing; Programs; Public Health Medicine (Field); Public Health Relevance; Scale Up; Screening; Screenings; Social Role; Tandem Mass Spectrometry; Technology Development