Enabling use of blood spot cards for accurate high throughput Fragile X screening

描述（由申请人提供）：本项目的长期目标是开发准确，快速，高通量和成本效益的筛查脆性X综合征（FXS）和相关疾病。FXS是自闭症和遗传性智力残疾最常见的已知遗传原因，大约每4,000名男性中有1名，每6,000名女性中有1名。脆性X染色体携带者的发病率在女性中高达1/130，在男性中高达1/250，并且这些个体存在影响儿童早期发育的其他疾病的风险，包括 自闭症新生儿筛查（NBS）脆性X是及时的，因为疾病的流行，缺乏一个明确的表型在出生时和相应的“诊断奥德赛”，目前和新兴的早期干预措施的选择，以及相关综合征的影响。然而，获得廉价且有效的高通量筛选技术一直是一个关键障碍。为了解决这一技术差距，Asuragen已经在I期研究中证明了用于血液卡处理和脆性X基因检测的精简和自动化系统的可行性，该系统利用了优化的PCR试剂组。第一阶段的研究非常成功，产生了一种可以一致和准确地检测脆性X细胞扩增的技术。在第二阶段的商业化中，这些能力将通过自动化一种新型的多重筛选试验，开发和整合一套创新的控制方法来扩展，以确保可靠的数据解释， 建立一个自动化的算法来识别阳性使用的工作流程，可以容纳每天数千个样本，并纳入确认方法，利用Asuragen的验证AmplideX”产品技术。因此，Asuragen将扩大以前成功的脆弱X SBIR提案，这些提案通过利用非常适合开发、优化、制造和商业化拟议的第二阶段产品的团队和环境，产生了多种尖端产品。本提案的具体目标是：目标1：在优化的工作流程中开发和整合一套质控品、标准品、试剂和QC指标，从而实现FMR 1正常和突变血点的高通量脆性X NBS。 目标二：建立一个整合自动化检测方法和软件的筛查系统，每天可自动分析多达4000个血斑样本。 目标3：在一项回顾性临床研究中使用该系统，该研究包括至少10，000份新生儿血斑样本，并对扩增的等位基因进行确认性反射测试。

DESCRIPTION (provided by applicant): The long term goal of this project is to develop accurate, rapid, high throughput and cost-effective screening for fragile X syndrome (FXS) and related disorders. FXS is the most common known genetic cause of autism and inherited intellectual disability, and affects roughly 1 in 4,000 males and 1 in 6,000 females. The incidence of fragile X carriers is as high as 1 in 130 in females and 1 in 250 in males, and these individuals are at risk for additional disorders that impact childhood early development, including autism. Newborn screening (NBS) for fragile X is timely due to the disease prevalence, lack of a clear phenotype at birth and corresponding "diagnostic odyssey", current and emerging options for early interventions, and impact of associated syndromes. Access to an inexpensive and effective high throughput screening technology, however, has been a key hurdle. To address this technology gap, Asuragen has demonstrated feasibility in phase I studies for a streamlined and automated system for blood card processing and fragile X gene detection that exploits an optimized PCR reagent set. Phase I studies were highly successful, resulting in a technology that can consistently and accurately detect fragile X expansions. In Phase II commercialization, these capabilities will be extended by automating a novel, multiplexed screening assay, developing and integrating an innovative set of controls that ensure reliable data interpretations, establishing an automated algorithm for identifying positives using a workflow that can accommodate thousands of samples per day, and incorporating a confirmation approach that leverages Asuragen's validated AmplideX" product technology. As a result, Asuragen will expand on previously successful fragile X SBIR proposals that have led to multiple cutting-edge products by leveraging a team and environment that is ideally suited to develop, optimize, manufacture, and commercialize the proposed Phase II product. The specific aims of this proposal are: Aim 1: Develop and integrate a set of controls, standards, reagents and QC metrics in an optimized workflow that enables high throughput fragile X NBS from FMR1 normal and mutant blood spots. Aim 2: Establish a screening system that integrates automated assay methods and software that can enable the automated analysis of up to 4000 blood spot samples per day. Aim 3: Validate the system in a retrospective clinical study that includes at least 10,000 newborn blood spot samples, with confirmatory reflex testing of expanded alleles.