MassARRAY基因分型技术在新生儿遗传代谢病诊断中的应用  

Application of MassARRAY genotyping assay in the diagnosis of neonatal genetic metabolic diseases

作　　者：张婷[1] 尚世强[2] 杨建滨[1] 周旭晨[3] 张玉 杨茹莱[1] 黄新文[1] Zhang Ting;Shang Shiqiang;Yang Jianbin;Zhou Xuchen;Zhang Yu;Yang Rulai;Huang Xinwen(Department of Genetics and Metabolism,the Children′s Hospital,Zhejiang University School of Medicine,National Clinical Research Center for Child Health,Hangzhou 310052;Lab Center,the Children′s Hospital,Zhejiang University School of Medicine,Hangzhou 310052,China;Department of Neonatology,the First Division Hospital of Xinjiang Production and Construction Corps,Aksu 843000,China;Zhejiang Biosan Biochemical Technologies Co.,Ltd,Hangzhou 310012,China)

机构地区：[1]浙江大学医学院附属儿童医院遗传与代谢科、国家儿童健康与疾病临床医学研究中心,杭州310052 [2]浙江大学医学院附属儿童医院实验检验中心,杭州310052 [3]新疆生产建设兵团第一师医院新生儿科,阿克苏843000 [4]浙江博圣生物技术股份有限公司,杭州310012

出　　处：《中华检验医学杂志》2023年第2期155-162,共8页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金(82073560);新疆生产建设兵团第一师医院精鹰计划(2021YN-JY-01)。

摘　　要：目的探讨MassARRAY基因分型技术应用于新生儿遗传代谢病诊断的准确性、时效性及可行性。方法回顾性研究。收集2016年12月至2020年1月在浙江省新生儿筛查中心应用串联质谱筛查的疑似阳性患儿7922例,采用MassARRAY技术进行27种遗传代谢病的常见变异位点检测,通过Sanger或二代测序验证并进一步寻找潜在变异。结果共1408份样本送检MassARRAY,307例确诊为遗传代谢病患儿,其中高苯丙氨酸血症检出率最高,其次为原发性肉碱缺乏症、短链酰基辅酶A脱氢酶缺乏症和甲基丙二酸血症。经Sanger测序验证100%(307/307)符合。287例检测为携带者,49.1%(141/287)经Sanger测序确认为携带者,以SLC22A5、MCCC1基因为主。50.8%(146/287)还检测到另一个等位基因变异,以PAH、PTS和ACADS基因为主。814例未发现变异,对其中158例复查后串联质谱特征性指标持续阳性、尿有机酸及其他生化检测等异常的样本进行二代测序,38%(60/158)检测到2个等位基因变异。最终确诊513例遗传代谢病患者,MassARRAY的总检出率为59.8%(307/513)。结论MassARRAY技术可作为新生儿遗传代谢病的早期分子筛查方法,在高苯丙氨酸血症、原发性肉碱缺乏症等热点变异集中的疾病中检出率较高,后期需不断优化新的疾病基因和变异位点从而进一步提升其潜在应用价值。Objective To investigate the accuracy,effectiveness and feasibility of MassARRAY genotyping assay in the diagnoses of neonatal genetic metabolic diseases.Methods This is a retrospective study.From December 2016 to January 2020,newborns were screened by tandem mass spectrometry at the Zhejiang Newborn Screening Center,among which the data of 7922 suspected positive cases of genetic metabolic diseases were collected.These patients were then tested for the common variants of 27 genetic metabolic diseases by MassARRAY genotyping assay,along with further testing using Sanger or next-generation sequencing used to verify and/or further search for potential variants.Results A total of 1408 cases were tested with MassARRAY.Among these,307 cases were confirmed with certain genetic metabolic diseases.The detection rate of hyperphenylalaninemia was the highest,followed by primary carnitine deficiency,short acyl-coA dehydrogenase deficiency and methylmalonic acidemia.With these cases,the consistency of Sanger sequencing and MassARRAY was 100%(307/307).Another 287 cases were identified as carriers by MassARRAY with a 49.1%(141/287)consistency in reference to Sanger sequencing,mainly involving SLC22A5 and MCCC1 genes.Meanwhile,50.8%(146/287)of these cases were found to have another variant mainly involving PAH,PTS and ACADS genes.The remaining 814 cases have no variants;158 cases out of these patients have continuously abnormal amino acids,acyl carnitines,urine organic acid and/or other biochemical indices,and were tested by next-generation sequencing,among which 38%(60/158)were detected with two variants.In this study,a total of 513 patients with genetic metabolic disease were diagnosed,and the detection rate of MassARRAY was 59.8%(307/513).Conclusions MassARRAY genotyping assay can be used as an early molecular screening method for neonatal genetic metabolic diseases.The detection rate is particularly high in diseases with a high concentration of hotspot variants,such as hyperphenylalaninemia and primary carnitine deficiency.

关 键 词：新生儿遗传代谢病 串联质谱法 基因分型技术 变异 应用价值 

分 类 号：R722.1[医药卫生—儿科]