Xu Zhong-Yao, Huang Shuo-Dan, Zou Jie, Zeng Wei-Hong, Guo Yong-Chao, Jiang Jian-Hui
Clin Lab. 2025 May 1;71(5). doi: 10.7754/Clin.Lab.2024.241006.
Next-generation sequencing (NGS) has greatly improved the diagnostic process for hereditary diseases, and incorporation of NGS into newborn screening (NBS) programs for more actionable diseases has been widely discussed. The aim of this study was to evaluate an integrated solution for application of NGS in newborn screening.
An NGS panel targeting 155 genes related to inborn errors of metabolism, hearing loss, severe combined immunodeficiency, congenital hypothyroidism, and other actionable genetic diseases, was designed. An all-in-one library preparation strategy was developed to combine multiplex PCR target enrichment and sample barcoding. A clinical genetic analysis system was assembled to facilitate bioinformatics analysis and reporting. The integrated solution was validated using 160 samples with known variants.
The end-to-end time from DNA isolation to sequencing was approximately 34 hours, and bioinformatics analysis pipeline took 4 hours for 160 samples in parallel. This allowed reporting of results on day 3. All known variants were confirmed by the NGS workflow, and two large insertion/deletions were additionally detected in two cases with previously clinically but not genetically confirmed diseases.
The integrated solution for application of NGS in NBS provided reasonable turnaround time to meet the NBS timeframe and could be implemented at scale.
下一代测序（NGS）极大地改善了遗传性疾病的诊断流程，并且将NGS纳入针对更多可采取行动疾病的新生儿筛查（NBS）项目已得到广泛讨论。本研究的目的是评估一种将NGS应用于新生儿筛查的综合解决方案。
设计了一个针对155个与先天性代谢缺陷、听力损失、重症联合免疫缺陷、先天性甲状腺功能减退及其他可采取行动的遗传疾病相关基因的NGS检测板。开发了一种一体化文库制备策略，将多重PCR目标富集和样本条形码技术相结合。组装了一个临床遗传分析系统以促进生物信息学分析和报告。使用160个已知变异的样本对该综合解决方案进行验证。
从DNA提取到测序的端到端时间约为34小时，生物信息学分析流程对160个样本并行处理需要4小时。这使得在第3天就能报告结果。所有已知变异均通过NGS工作流程得到确认，并且在两例之前临床诊断但基因未确诊的疾病病例中额外检测到两个大的插入/缺失。
将NGS应用于新生儿筛查的综合解决方案提供了合理的周转时间以满足新生儿筛查的时间框架，并且可以大规模实施。
