PDF(Pubmed)
Abstract：
BACKGROUND: Newborn screening (NBS), such as tandem mass spectrometry (MS/MS), may yield false positive/negative results. Next-generation sequencing (NGS) has the potential to provide increased data output, efficiencies, and applications. This study aimed to analyze the types and distribution of pathogenic gene mutations in newborns in Huzhou, Zhejiang province, China and explore the applicability of NGS and MS/MS in NBS.
METHODS: Blood spot samples from 1263 newborns were collected. NGS was employed to screen for pathogenic variants in 542 disease-causing genes, and detected variants were validated using Sanger sequencing. Simultaneously, 26 inherited metabolic diseases (IMD) were screened using MS/MS. Positive or suspicious samples identified through MS/MS were cross-referenced with the results of NGS.
RESULTS: Among all newborns, 328 had no gene mutations detected. NGS revealed at least one gene mutation in 935 newborns, with a mutation rate of 74.0%. The top 5 genes were FLG, GJB2, UGT1A1, USH2A, and DUOX2. According to American College of Medical Genetics guidelines, gene mutations in 260 cases were classified as pathogenic or likely pathogenic mutation, with a positive rate of 20.6%. The top 5 genes were UGT1A1, FLG, GJB2, MEFV, and G6PD. MS/MS identified 18 positive or suspicious samples for IMD and 1245 negative samples. Verification of these cases by NGS results showed no pathogenic mutations, resulting in a false positive rate of 1.4% (18/1263).
CONCLUSIONS: NBS using NGS technology broadened the range of diseases screened, and enhanced the accuracy of diagnoses in comparison to MS/MS for screening IMD. Combining NGS and biochemical screening would improve the efficiency of current NBS.
METHODS: Blood spot samples from 1263 newborns were collected. NGS was employed to screen for pathogenic variants in 542 disease-causing genes, and detected variants were validated using Sanger sequencing. Simultaneously, 26 inherited metabolic diseases (IMD) were screened using MS/MS. Positive or suspicious samples identified through MS/MS were cross-referenced with the results of NGS.
RESULTS: Among all newborns, 328 had no gene mutations detected. NGS revealed at least one gene mutation in 935 newborns, with a mutation rate of 74.0%. The top 5 genes were FLG, GJB2, UGT1A1, USH2A, and DUOX2. According to American College of Medical Genetics guidelines, gene mutations in 260 cases were classified as pathogenic or likely pathogenic mutation, with a positive rate of 20.6%. The top 5 genes were UGT1A1, FLG, GJB2, MEFV, and G6PD. MS/MS identified 18 positive or suspicious samples for IMD and 1245 negative samples. Verification of these cases by NGS results showed no pathogenic mutations, resulting in a false positive rate of 1.4% (18/1263).
CONCLUSIONS: NBS using NGS technology broadened the range of diseases screened, and enhanced the accuracy of diagnoses in comparison to MS/MS for screening IMD. Combining NGS and biochemical screening would improve the efficiency of current NBS.
摘要：
背景：新生儿筛查（NBS），如串联质谱(MS/MS)，可能会产生假阳性/阴性结果。下一代测序（NGS）有可能提供更多的数据输出，效率，和应用。本研究旨在分析湖州市新生儿致病基因突变的类型和分布,浙江省,并探讨了NGS和MS/MS在NBS中的适用性。
方法：收集1263例新生儿的血斑样本。NGS被用来筛选542个致病基因中的致病变异，并使用Sanger测序验证检测到的变异。同时,使用MS/MS筛选了26种遗传代谢疾病（IMD）。通过MS/MS鉴定的阳性或可疑样品与NGS的结果交叉参考。
结果：在所有新生儿中，328没有检测到基因突变。NGS揭示了935名新生儿中至少有一个基因突变，突变率为74.0%。前5个基因是FLG,GJB2,UGT1A1,USH2A,DUOX2根据美国医学遗传学学会的指南,260例基因突变被分类为致病性或可能的致病性突变，阳性率为20.6%。前5位基因分别为UGT1A1、FLG、GJB2,MEFV,G6PDMS/MS鉴定出18个IMD阳性或可疑样品和1245个阴性样品。通过NGS结果对这些病例的验证显示没有致病性突变，导致假阳性率为1.4%(18/1263)。
结论：使用NGS技术的NBS扩大了筛查疾病的范围，与MS/MS筛查IMD相比，提高了诊断的准确性。结合NGS和生化筛选将提高当前NBS的效率。
方法：收集1263例新生儿的血斑样本。NGS被用来筛选542个致病基因中的致病变异，并使用Sanger测序验证检测到的变异。同时,使用MS/MS筛选了26种遗传代谢疾病（IMD）。通过MS/MS鉴定的阳性或可疑样品与NGS的结果交叉参考。
结果：在所有新生儿中，328没有检测到基因突变。NGS揭示了935名新生儿中至少有一个基因突变，突变率为74.0%。前5个基因是FLG,GJB2,UGT1A1,USH2A,DUOX2根据美国医学遗传学学会的指南,260例基因突变被分类为致病性或可能的致病性突变，阳性率为20.6%。前5位基因分别为UGT1A1、FLG、GJB2,MEFV,G6PDMS/MS鉴定出18个IMD阳性或可疑样品和1245个阴性样品。通过NGS结果对这些病例的验证显示没有致病性突变，导致假阳性率为1.4%(18/1263)。
结论：使用NGS技术的NBS扩大了筛查疾病的范围，与MS/MS筛查IMD相比，提高了诊断的准确性。结合NGS和生化筛选将提高当前NBS的效率。
