目的 评估二代测序在新生儿疾病筛查中的临床应用价值，尤其是与串联质谱法联合应用的优势。 方法 前瞻性收集2021年5—8月于深圳市光明区人民医院出生的1 999例新生儿的血样，所有样本均应用串联质谱和荧光免疫分析法初筛，采用二代测序检测135个高频致病基因变异位点，并对可疑阳性位点应用Sanger测序或多重连接探针扩增技术进行家系验证。 结果 1 999例新生儿的串联质谱分析未发现确诊阳性患儿。基因筛查发现58例阳性（2.90%），732例为致病基因携带（36.62%），1 209例阴性（60.48%）；确诊新生儿肝内胆汁淤积症1例（0.05%，1/1 999）。荧光免疫分析发现葡萄糖-6-磷酸脱氢酶（glucose-6-phosphate dehydrogenase, G6PD）缺乏症39例（1.95%，39/1 999），基因筛查发现G6PD缺乏症43例（2.15%，43/1 999）。荧光免疫分析筛查发现高促甲状腺激素血症患儿6例（0.30%，6/1 999），基因筛查结果均携带DUOX2基因变异。致病基因携带率前10位的基因为G6PD（12.8%）、DUOX2（8.7%）、HBB（8.2%）、ATP7B（6.6%）、GJB2（5.7%）SLC26A4（5.6%）、PAH（5.6%）ACADSB（4.6%）、SLC25A13（4.2%）、SLC22A5（4.1%）。 结论 二代测序可作为串联质谱法的有效补充，可显著提高新生儿遗传代谢病的检出率，结合家系验证能实现精准诊断，尤其对G6PD缺乏症等单基因疾病的筛查具有互补优势。
Abstract
Objective To evaluate the clinical value of next-generation sequencing (NGS) in neonatal disease screening, particularly its advantages when combined with tandem mass spectrometry (MS/MS). Methods A prospective study was conducted involving blood samples from 1 999 neonates born at the Shenzhen Guangming District People's Hospital, between May and August 2021. All samples were initially screened using MS/MS and fluorescence immunoassay, followed by NGS to detect high-frequency variation sites in 135 related pathogenic genes. Suspected positive variants were validated using Sanger sequencing or multiplex ligation-dependent probe amplification in family studies. Results No confirmed positive cases were found in the MS/MS analysis of the 1 999 neonates. Genetic screening identified 58 positive cases (2.90%), 732 carriers of pathogenic genes (36.62%), and 1 209 negative cases (60.48%). One case of neonatal intrahepatic cholestasis was diagnosed (0.05%, 1/1 999). Fluorescence immunoassay identified 39 cases of glucose-6-phosphate dehydrogenase (G6PD) deficiency (1.95%, 39/1 999), while genetic screening identified 43 cases of G6PD deficiency (2.15%, 43/1 999). The fluorescence immunoassay also detected 6 cases of hyperthyrotropinemia (0.30%, 6/1 999), all of whom carried DUOX2 gene variants. The top ten pathogenic gene carrier rates were G6PD (12.8%), DUOX2 (8.7%), HBB (8.2%), ATP7B (6.6%), GJB2 (5.7%), SLC26A4 (5.6%), PAH (5.6%), ACADSB (4.6%), SLC25A13 (4.2%), and SLC22A5 (4.1%). Conclusions NGS can serve as an effective complement to MS/MS, significantly improving the detection rate of inherited metabolic disorders in neonates. When combined with family validation, it enables precise diagnosis, particularly demonstrating complementary advantages in screening for monogenic diseases such as G6PD deficiency.
关键词
基因筛查 /
串联质谱 /
新生儿筛查 /
遗传代谢病 /
新生儿
Key words
Genetic screening /
Tandem mass spectrometry /
Neonatal screening /
Inherited metabolic disorder /
Neonate
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