Targeted next generation sequencing for newborn screening of Menkes disease
Targeted next generation sequencing for newborn screening of Menkes disease
复制标题
DOI：
10.1016/j.ymgmr.2020.100625
复制
发表时间：
2020-09-01
影响因子：
1.9
通讯作者：
Bhattacharjee, Arindam
中科院分区：
文献类型：
--
作者：
Parad, Richard B.;Kaler, Stephen G.;Bhattacharjee, Arindam
目的：基于人群的新生儿筛查（NBS）可以早期发现和治疗遗传性疾病。然而，对于某些医学上可行的病症，NBS受到缺乏可靠的生物化学特征的限制，该生物化学特征适于通过当前平台检测。我们试图评估的分析有效性的ATP 7A为目标的下一代DNA测序检测作为一个潜在的新生儿屏幕为这样的疾病，Menkes diseases.Methods：从控制或Menkes病受试者（n = 22）的干血斑进行了盲分析的致病性变异的铜转运基因，ATP 7A。该分析方法进行了优化，以最大限度地降低成本，并提供快速的周转时间。结果：该算法正确识别致病性ATP 7A变异，包括错义，无义，小插入/缺失，和大拷贝数的变异，在21/22（95.5%）的主题，其中一个人以前有不确定的诊断测序。对于一个假阴性，也没有被检测到的商业分子实验室，我们确定了一个深内含子变异，损害ATP 7A mRNA splicing.Conclusions：我们的研究结果支持的概念证明，主要基于DNA的NBS将准确地检测门克斯病，一种疾病，满足威尔逊和Jungner筛选标准，生化NBS不可用。针对NBS的下一代靶向测序将能够改善Menkes病的临床结果，建立一个早期识别其他未筛查疾病的平台，并通过为许多生化筛查条件的分子确认提供即时数据来补充当前的NBS。
Purpose: 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 ATP7A targeted next generation DNA sequencing assay as a potential newborn screen for one such disorder, Menkes disease.Methods: Dried blood spots from control or Menkes disease subjects (n = 22) were blindly analyzed for pathogenic variants in the copper transport gene, ATP7A. The analytical method was optimized to minimize cost and provide rapid turnaround time.Results: The algorithm correctly identified pathogenic ATP7A 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 ATP7A mRNA splicing.Conclusions: 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 conditions.