An automated microfluidic platform for high-throughput newborn SCID screening

用于高通量新生儿 SCID 筛查的自动化微流控平台

• 批准号: 8058161
• 负责人: Vijay Srinivasan
• 金额: $ 17.97万
• 依托单位: ADVANCED LIQUID LOGIC
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8058161
• 关键词: Actins; Address; Adopted; Adoption; Advisory Committees; Affect; Automation; Biological Assay; Birth; Blood; Bone Marrow Transplantation; Budgets; Child; Collaborations; Color; DNA; DNA Microarray Chip; DNA analysis; Detection; Development; Diagnosis; Discrimination; Disease; Early Diagnosis; Early treatment; Economics; Equipment; Family history of; Genes; Goals; Gold; Grant; Human Resources; Illinois; Image; Immune; Infant; Infection; Investments; Laboratories; Lead; Life; Liquid substance; Maintenance; Massachusetts; Methodology; Methods; Microfluidics; Molecular; Monitor; Neonatal Screening; Newborn Infant; Optics; Phase; Physicians; Preparation; Price; Process; Protocols documentation; Public Health; Reaction; Recommendation; Research; Robotics; Sampling; Screening procedure; Severe Combined Immunodeficiency; Solutions; Speed; Spottings; Survival Rate; System; T-Cell Development; T-Cell Receptor-Rearrangement Excision DNA Circles; Technology; Temperature; Testing; Time; Training; Transplantation; Wisconsin; base; boys; cost; cost effective; design; digital; infancy; infant death; instrument; instrumentation; internal control; micro-total analysis system; pilot trial; population based; programs; reconstitution

DESCRIPTION (provided by applicant): 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.

描述（由申请人提供）：这项研究合作的总体目标是开发一种高通量数字微流体仪器，能够同时筛查数十名新生儿的严重联合免疫缺陷（SCID）。鉴于2010年1月21日，新生儿和儿童遗传性疾病联邦咨询委员会（ACHDNC）一致建议将严重联合免疫缺陷（SCID）添加到统一新生儿筛查小组中，因此在州筛查水平上非常需要这样的技术平台。为统一新生儿筛查小组增加的条件成为各州采用筛查计划的强大动力，因此，低成本、高通量的解决方案是面临严重预算危机的州的关键。除非能够实现免疫重建，否则SCID通常在生命的第一年或第二年是致命的。众所周知，骨髓移植导致在3.5个月前接受这种治疗的人的存活率为94%。SCID婴儿在出生时或婴儿早期没有外在的身体异常来提醒医生存在这种情况，因此通常只有在严重感染发展后才能做出诊断。很明显，新生儿筛查是早期诊断和最佳治疗大多数患有这种疾病的婴儿的唯一希望。目前，新生儿筛查的方法是从婴儿身上收集干血斑，然后送到实验室进行分析。虽然成本估计从过去的威斯康星州试点新生儿SCID筛查计划估计每测试5 - 6美元，仍然有许多前期费用与实施这项测试。例如，TREC检测涉及许多DNA提取步骤，需要对人员进行高级培训。同样，高通量实时PCR仪器需要昂贵的前期和维护成本。随后，我们坚信，用于在新生儿筛查中执行SCID DNA分析测定的数字微流体平台将以非常低的成本实现自动化。在这个项目中，将开发一个完整的TREC提取和检测数字微流控平台，没有移动部件，用于3mm打孔干燥血点。所有提取、纯化和浓缩DNA、对TREC进行后续热循环PCR以及光学检测的步骤都将围绕ALL的核心数字微流体平台开发。我们将使用约100份正常干血斑样本和约10份白细胞减少样本（代表SCID样本）进行验证，以证明正常和SCID受影响的血斑之间的区分。在完成这项资助的拟议目标后，我们相信我们将接近于拥有一个完全自动化的高通量解决方案，不仅可以用于国家实验室的SCID筛查，还可以用于其他新生儿疾病。 公共卫生相关性：严重的联合免疫缺陷（更普遍地称为“泡泡男孩病”）在生命的第一年或第二年是普遍致命的。骨髓移植可以为这种疾病带来挽救生命的免疫重建，在出生后3.5个月内接受移植的婴儿的存活率为96%。该提案的中心目标是开发一个完整的高通量数字微流控DNA分析芯片实验室平台，这将导致一个非常便宜和完全自动化的解决方案，用于筛选新生儿这种致命的条件。这将降低公共卫生实验室进行这种疾病筛查的技术和财政障碍。