Lab-on-a-Chip for Multiplexed Newborn Screening of Lysosomal Storage Disorders

描述(申请人提供)：目前进行新生儿筛查的方法是从婴儿身上采集干燥的血迹，然后将其送往实验室进行分析。它主要是为了检测导致严重身体和精神残疾的障碍。越来越有必要对一些溶酶体储存障碍进行筛查，治疗方法正在变得可用。串联质谱仪是一种广泛应用于新生儿筛查的多重检测技术，但将其应用于酶分析存在成本高、耗时长、劳动强度大等缺点。需要一种廉价、快速和自动化的新生儿筛查分析技术。一种数字微流体盒，本质上是在亚微升液滴上操作的液体处理系统，具有 已开发用于使用干血斑进行用于筛选溶酶体储存障碍的荧光酶分析。该系统被配置为能够实现自动操作并廉价地多路传输多个化验。在第二阶段，我们已经成功地演示了Pompe、Fabry、Gaucher、Hunter和Hurler疾病的分析方法，只需从干血点上一击即可。在所有5种疾病中，数字微流控方法与杜克大学在微量平板上进行的“金标准”荧光酶分析方法相比都是有利的。数字微流控系统目前正在伊利诺伊州的新生儿中进行试点试验 筛查实验室。在正在进行的试点研究中，已经对6000多个干血点进行了筛查，以确定庞佩、法布里和高歇尔的情况。在该平台上获得阳性结果后，4名婴儿进一步确诊为法布里病，1名婴儿确诊为高谢病。第二阶段B的继续工作将侧重于解决在试点试验期间发现的几个用户问题，向FDA提交510(K)申请，并扩大新生儿筛查化验小组，以包括其他已获授权或已有治疗方法/正在开发的LSD。 与公共健康相关：几乎每个在美国出生的婴儿都要接受新生儿筛查。由于治疗方法的可获得性，人们对将溶酶体储存障碍(LSD)添加到测试小组中的兴趣越来越大。在这个项目中，将开发和验证一个具有成本效益的微流控平台，用于新生儿LSD的筛查。伊利诺伊州、密苏里州和新墨西哥州已制定法律，要求对所有婴儿进行LSD筛查，这种廉价平台的提供将使资金紧张的州负担得起。在伊利诺伊州对该平台进行有限的试点试验期间，一些患有LSD的婴儿已经被识别出来。

DESCRIPTION (provided by applicant): Newborn screening is currently performed by collecting dried bloodspots from infants and then sending them to a lab for analysis. It is primarily performed to detect disorders which lead to severe physical and mental disabilities. There is an increasing necessity to screen for a number of lysosomal storage disorders for which therapies are becoming available. Tandem mass spectrometry is a multiplex detection technology widely utilized in newborn screening, but when applied to enzyme assays it is expensive, time consuming, and labor intensive. There is a need for an inexpensive, rapid, and automated technology for performing newborn screening assays. A digital microfluidic cartridge, which is essentially a liquid handling system operating on sub- microliter droplets, has been developed for performing fluorometric enzymatic assays for screening lysosomal storage disorders using dried blood spots. This system is configured to enable walkaway automation and multiplex several assays inexpensively. In phase II, we have successfully demonstrated assays for Pompe, Fabry, Gaucher, Hunter and Hurler disorders using a single punch from a dried blood spot. The digital microfluidic method compared favorably with a "gold standard" fluorometric enzymatic method performed in microtiter plates at Duke University for all the 5 disorders. The digital microfluidic system is currently under a pilot trial in the Illinois Newborn Screening Laboratory. Over 6,000 dried blood spots have been screened for Pompe, Fabry, and Gaucher during the ongoing pilot study. Following a positive results on this platform, 4 infants were further confirmed with Fabry disease and 1 infant with Gaucher disease. Phase IIB continuation will focus on addressing several user issues that were identified during the pilot trial, submission of a 510(k) application to the FDA, and expansion of the newborn screening assay panel to include other LSDs which are mandated or which have therapies available/under development. PUBLIC HEALTH RELEVANCE: Newborn screening is performed on almost every infant born in the US. Due to the availability of therapies, there is a growing interest in adding lysosomal storage disorders (LSDs) to the panel of tests. In this project, a cost-effective microfluidic platform will be developed and validated for newborn screening of LSDs. Illinois, Missouri and New Mexico have mandated laws requiring screening all the babies for LSDs and availability of this inexpensive platform will make it affordable for cash-strapped states. During a limited pilot trial of this platform in Illinois, some babies with LSDs were already identified.