Parad Richard B, Kaler Stephen G, Mauceli Evan, Sokolsky Tanya, Yi Ling, Bhattacharjee Arindam
Department of Pediatric Newborn Medicine, Brigham & Women's Hospital, Harvard Medical School, Boston, MA, United States of America.
Section on Translational Neuroscience, Molecular Medicine Branch, Eunice Kennedy Shriver National Institute of Child Health and Human Development, Bethesda, MD, United States of America.
Mol Genet Metab Rep. 2020 Jul 21;24:100625. doi: 10.1016/j.ymgmr.2020.100625. eCollection 2020 Sep.
Population-based newborn screening (NBS) allows early detection and treatment of inherited disorders. For certain medically-actionable conditions, however, NBS is limited by the absence of reliable biochemical signatures amenable to detection by current platforms. We sought to assess the analytic validity of an targeted next generation DNA sequencing assay as a potential newborn screen for one such disorder, Menkes disease.
Dried blood spots from control or Menkes disease subjects ( = 22) were blindly analyzed for pathogenic variants in the copper transport gene, The analytical method was optimized to minimize cost and provide rapid turnaround time
The algorithm correctly identified pathogenic variants, including missense, nonsense, small insertions/deletions, and large copy number variants, in 21/22 (95.5%) of subjects, one of whom had inconclusive diagnostic sequencing previously. For one false negative that also had not been detected by commercial molecular laboratories, we identified a deep intronic variant that impaired mRNA splicing.
Our results support proof-of-concept that primary DNA-based NBS would accurately detect Menkes disease, a disorder that fulfills Wilson and Jungner screening criteria and for which biochemical NBS is unavailable. Targeted next generation sequencing for NBS would enable improved Menkes disease clinical outcomes, establish a platform for early identification of other unscreened disorders, and complement current NBS by providing immediate data for molecular confirmation of numerous biochemically screened condition.
基于人群的新生儿筛查（NBS）可实现对遗传性疾病的早期检测和治疗。然而，对于某些可采取医学行动的疾病，NBS受到当前平台无法检测到可靠生化特征的限制。我们试图评估一种靶向新一代DNA测序检测方法作为一种潜在的新生儿筛查方法用于一种此类疾病——门克斯病的分析有效性。
对来自对照或门克斯病患者（n = 22）的干血斑进行盲法分析，以检测铜转运基因中的致病变异。对分析方法进行了优化，以尽量降低成本并缩短周转时间。
该算法在21/22（95.5%）的受试者中正确识别出了致病变异，包括错义、无义、小插入/缺失和大拷贝数变异，其中一名受试者此前诊断性测序结果不明确。对于一个商业分子实验室也未检测到的假阴性结果，我们鉴定出了一个影响mRNA剪接的内含子深处变异。
我们的结果支持基于DNA的原发性NBS能够准确检测出门克斯病的概念验证，门克斯病符合威尔逊和荣格纳筛查标准且目前尚无生化NBS方法。用于NBS的靶向新一代测序将改善门克斯病的临床结局，建立一个早期识别其他未筛查疾病的平台，并通过为众多生化筛查疾病的分子确认提供即时数据来补充当前的NBS。
