The overall goal of this research collaboration is to develop a high-throughput digital microfluidic instrument capable of screening dozens of newborns simultaneously for severe combined immunodeficiency (SCID). There is a significant need at the state-screening level for such a technology platform given that on January 21, 2010, a federal Advisory Committee on Heritable Disorders in Newborns and Children (ACHDNC), unanimously recommended that severe combined immunodeficiency (SCID) be added to the Uniform Newborn Screening Panel. Conditions added to the Uniform Newborn Screening Panel become a strong motivator for states to adopt screening programs, so a low-cost, high-throughput solution is a key for states facing critical budget crises. SCID is universally fatal in the first or second year of life unless immune reconstitution can be achieved. It is known that bone marrow transplantation results in a 94% survival rate for those given this treatment before 3.5 months of life. SCID infants have no outward physical abnormalities at birth or in early infancy to alert physicians that the condition exists, so the diagnosis is usually made only after serious infections develop. It is clear that newborn screening is the only hope for early diagnosis and optimal treatment for most infants with this condition. Newborn screening is currently performed by collecting dried bloodspots from infants and then sending them to a lab for analysis. While cost estimates from a past Wisconsin pilot newborn SCID screening program estimated $5-6 per test, there are still many upfront costs associated with implementing this test. For instance TREC detection involves many DNA extraction steps requiring advanced training of personnel. Likewise, high-throughput real-time PCR instrumentation requires costly upfront and maintenance costs. Subsequently, we strongly believe that a digital microfluidic platform for performing DNA analysis assays for SCID in newborn screening will enable walkaway automation at a very low cost. In this project, a complete TREC extraction and detection digital microfluidic platform with no moving parts will be developed for use with 3mm punch dried blood spots. All the steps for extracting, purifying, and concentrating DNA, performing subsequent thermocycled PCR for TRECs, and optical detection will be developed around ALL's core digital microfluidic platform. We will validate with about 100 normal dried blood spot samples and about 10 leukoreduced samples (representing SCID samples) to demonstrate discrimination between normal and SCID affected spots. Upon completion of the proposed aims in this grant, we believe we will be near to a having a completely automated high-throughput solution that can not only be leveraged for SCID screening in the state labs, but for other newborn disorders, as well.
Public Health Relevance: Severe combined immunodeficiency (more popularly known as "bubble boy disease") is universally fatal in the first or second year of life. Bone marrow transplantation can result in life-saving immune reconstitution for this condition with a 96% survival rate for the infants that undergo transplantation before 3.5 months of life. The central goal of this proposal is to develop a complete high-throughput, digital microfluidic DNA analysis lab-on- a-chip platform that would lead to a significantly inexpensive and completely automated solution for screening newborns for this lethal condition. This would lower the technology and financial barrier for public health laboratories to undertake screening for this condition.
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