NEWBORN SCREENING BY MULTIPLEX MOLECULAR ANALYSIS

新生儿筛查是一项行之有效的公共卫生服务。串联质谱仪等创新方法大大增加了新生儿筛查实验室有效识别的疾病数量。为了继续扩大在新生儿中发现的可治疗疾病的数量，将开发一种第一级的、自动化的、多重分子检测。这项检测是基于从全球收集的新生儿血卡中获得的DNA。我们将建立多重聚合酶链式反应方法来扩增可诊断以下疾病的基因序列：镰状细胞血红蛋白病S、C和E等位基因；α-1-抗胰蛋白酶缺乏症S等位基因和Z等位基因；因子V莱顿A1691G；以及血色沉着症G845A。将使用低密度寡核苷酸阵列对聚合酶链式反应产物进行分析。低密度寡核苷酸阵列将同时区分所述基因座上的野生型和突变型等位基因。来自阵列的数据将使用BioScan近端成像系统进行处理。完成这项提案的具体目标将证明第一级分子筛查方案的可行性。同时检测几种疾病的能力将使全面的新生儿筛查更负担得起，并使公共卫生服务更有效。建议的商业应用：Neo Gen Screen提供了一种补充的筛查服务，超出了国家规定的范围。扩大发现的可治疗疾病的数量将使这项服务更加全面，从而对卫生保健机构更具吸引力。拟议的筛查服务适用于新生儿筛查和高危筛查。

Neonatal screening is a proven public health service. Innovative methods such as tandem mass spectrometry have greatly increased the number of disorders effectively identified in newborn screening laboratories. To continue expanding the number of treatable disorders identified in newborns, a first tier, automated, multiplex molecular assay will be developed. The assay is based on DNA obtained from the universally collected neonatal blood card. A multiplex PCR assay will be developed to amplify DNA sequences that are diagnostic for the following disorders: sickle cell hemoglobinopathies S, C, and E alleles; alpha-1-Antitrypsin Deficiency S allele and Z allele; Factor V Leiden A1691G; and Hemochromatosis G845A. Analysis of PCR products will be performed using a low-density oligonucleotide array. The low-density oligonucleotide array will simultaneously distinguish both wild type and mutant alleles at the loci described. Data from the arrays will be processed using a BioScan Proximal Imaging System. Completing the Specific Aims of this proposal will demonstrate the feasibility of a first tier molecular screening protocol. The ability to simultaneously detect several disorders will make comprehensive newborn screening more affordable and a more effective public health service. PROPOSED COMMERCIAL APPLICATIONS: Neo Gen Screening provides a supplemental screening service, beyond that mandated by the state. Expanding the number of treatable disorders detected will make the service more comprehensive and thus more attractive to health care institutions. The screening services proposed are marketable for newborn screening and high-risk screening.