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

描述(由申请人提供)：该项目的长期目标是开发准确、快速、高通量和成本效益高的脆性X综合征(FXS)及相关疾病筛查。FXS是导致自闭症和遗传性智力障碍的最常见的遗传原因，大约每4000名男性和每6000名女性中就有一人受到影响。脆性X携带者的发病率在女性中高达每130人中有1人，在男性中高达1/250，这些人面临着影响儿童早期发育的额外疾病的风险，包括 自闭症。新生儿脆性X筛查(NBS)是及时的，因为疾病流行，出生时缺乏明确的表型和相应的“诊断奥德赛”，当前和正在出现的早期干预选择，以及相关综合征的影响。然而，获得廉价有效的高通量筛查技术一直是一个关键障碍。为了解决这一技术差距，Asuragen已经在第一阶段研究中证明了利用优化的PCR试剂集进行血卡处理和脆性X基因检测的简化和自动化系统的可行性。第一阶段的研究非常成功，产生了一种可以持续和准确地检测脆性X膨胀的技术。在第二阶段的商业化中，这些能力将通过自动化新的、多路筛选分析、开发和整合一套确保可靠数据解释的创新控制来扩展， 使用每天可容纳数千个样本的工作流程建立用于确定阳性结果的自动化算法，并采用利用Asuragen经验证的AmplideX“产品技术的确认方法。因此，Asuragen将通过利用一个非常适合开发、优化、制造和商业化拟议的第二阶段产品的团队和环境，来扩展之前成功的Fragile X SBIR提议，这些提议已经导致了多种尖端产品。这项建议的具体目标是：目标1：在优化的工作流程中开发和集成一套控制、标准、试剂和质量控制指标，以实现来自FMR1正常和突变血点的高吞吐量脆性X NBS。目标2：建立一个集自动化化验方法和软件于一体的筛查系统，每天可以自动分析多达4000个血斑样本。目的3：在一项包括至少10,000份新生儿血斑样本的回顾性临床研究中验证该系统，并对扩展的等位基因进行确认性反射试验。

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.