NEWBORN SCREENING BY MULTIPLEX MOLECULAR ANALYSIS

新生儿筛查是一项经过验证的公共卫生服务。诸如串联质谱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、自动化的多重分子检测方法。该分析是基于从普遍收集的新生儿血液卡中获得的DNA。将开发多重PCR检测来扩增用于诊断以下疾病的DNA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；因素V莱顿A1691G；血色素沉着病G845A。PCR产物的分析将使用低密度寡核苷酸阵列进行。低密度寡核苷酸阵列将同时在所描述的位点上区分野生型和突变等位基因。来自阵列的数据将使用biscan近端成像系统进行处理。完成本提案的具体目标将证明第一层分子筛选方案的可行性。同时发现几种疾病的能力将使全面的新生儿筛查更便宜，并成为更有效的公共卫生服务。拟议的商业应用：Neo Gen筛选提供了一项补充筛选服务，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，从而对保健机构更具吸引力。建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.