Abstract：
Newborn screening (NBS) has been implemented for neonatal inborn disorders using various technology platforms, but false-positive and false-negative results are still common. In addition, target diseases of NBS are limited by suitable biomarkers. Here we sought to assess the feasibility of further improving the screening using next-generation sequencing technology.
We designed a newborn genetic sequencing (NBGS) panel based on multiplex PCR and next generation sequencing to analyze 134 genes of 74 inborn disorders, that were validated in 287 samples with previously known mutations. A retrospective cohort of 4986 newborns was analyzed and compared with the biochemical results to evaluate the performance of this panel.
The accuracy of the panel was 99.65% with all samples, and 154 mutations from 287 samples were 100% detected. In 4986 newborns, a total of 113 newborns were detected with biallelic or hemizygous mutations, of which 36 newborns were positive for the same disorder by both NBGS and conventional NBS (C-NBS) and 77 individuals were NBGS positive/C-NBS negative. Importantly, 4 of the 77 newborns were diagnosed currently including 1 newborn with methylmalonic acidemia, 1 newborn with primary systemic carnitine deficiency and 2 newborns with Wilson\'s disease. A total of 1326 newborns were found to be carriers with an overall carrier rate of 26.6%.
Analysis based on next generation sequencing could effectively identify neonates affected with more congenital disorders. Combined with C-NBS, this approach may improve the early and accurate identification of neonates with inborn disorders. Our study lays the foundation for prospective studies and for implementing NGS-based analysis in NBS.
We designed a newborn genetic sequencing (NBGS) panel based on multiplex PCR and next generation sequencing to analyze 134 genes of 74 inborn disorders, that were validated in 287 samples with previously known mutations. A retrospective cohort of 4986 newborns was analyzed and compared with the biochemical results to evaluate the performance of this panel.
The accuracy of the panel was 99.65% with all samples, and 154 mutations from 287 samples were 100% detected. In 4986 newborns, a total of 113 newborns were detected with biallelic or hemizygous mutations, of which 36 newborns were positive for the same disorder by both NBGS and conventional NBS (C-NBS) and 77 individuals were NBGS positive/C-NBS negative. Importantly, 4 of the 77 newborns were diagnosed currently including 1 newborn with methylmalonic acidemia, 1 newborn with primary systemic carnitine deficiency and 2 newborns with Wilson\'s disease. A total of 1326 newborns were found to be carriers with an overall carrier rate of 26.6%.
Analysis based on next generation sequencing could effectively identify neonates affected with more congenital disorders. Combined with C-NBS, this approach may improve the early and accurate identification of neonates with inborn disorders. Our study lays the foundation for prospective studies and for implementing NGS-based analysis in NBS.
摘要：
新生儿筛查（NBS）已使用各种技术平台对新生儿先天性疾病实施，但是假阳性和假阴性结果仍然很常见。此外,NBS的靶疾病受到合适的生物标志物的限制。在这里，我们试图评估使用下一代测序技术进一步改进筛查的可行性。
我们设计了一个基于多重PCR和下一代测序的新生儿基因测序（NBGS）小组，以分析74种先天性疾病的134个基因。在287个具有先前已知突变的样本中进行了验证。对4986例新生儿的回顾性队列进行了分析，并与生化结果进行了比较，以评估该小组的表现。
所有样品的面板精度为99.65％，从287个样本中检测到154个突变100%。在4986名新生儿中，共有113例新生儿检测到双等位基因或半合子突变，其中36例新生儿在NBGS和常规NBS（C-NBS）中均为同一疾病阳性，77例NBGS阳性/C-NBS阴性。重要的是,77名新生儿中有4名被诊断为目前包括1名患有甲基丙二酸血症的新生儿，1例新生儿原发性全身肉碱缺乏症和2例新生儿威尔逊病。共发现1326名新生儿为携带者，总携带者率为26.6%。
基于下一代测序的分析可以有效地识别患有更多先天性疾病的新生儿。结合C-NBS，这种方法可以提高新生儿先天性疾病的早期和准确识别。我们的研究为前瞻性研究和在NBS中实施基于NGS的分析奠定了基础。
我们设计了一个基于多重PCR和下一代测序的新生儿基因测序（NBGS）小组，以分析74种先天性疾病的134个基因。在287个具有先前已知突变的样本中进行了验证。对4986例新生儿的回顾性队列进行了分析，并与生化结果进行了比较，以评估该小组的表现。
所有样品的面板精度为99.65％，从287个样本中检测到154个突变100%。在4986名新生儿中，共有113例新生儿检测到双等位基因或半合子突变，其中36例新生儿在NBGS和常规NBS（C-NBS）中均为同一疾病阳性，77例NBGS阳性/C-NBS阴性。重要的是,77名新生儿中有4名被诊断为目前包括1名患有甲基丙二酸血症的新生儿，1例新生儿原发性全身肉碱缺乏症和2例新生儿威尔逊病。共发现1326名新生儿为携带者，总携带者率为26.6%。
基于下一代测序的分析可以有效地识别患有更多先天性疾病的新生儿。结合C-NBS，这种方法可以提高新生儿先天性疾病的早期和准确识别。我们的研究为前瞻性研究和在NBS中实施基于NGS的分析奠定了基础。
