石家庄地区枫糖尿病患儿串联质谱筛查及BCKDHA、BCKDHB、DBT基因突变分析

doi:10.12280/gjszjk.20230033

摘要/Abstract

摘要：

目的： 了解中国河北省石家庄地区新生儿中枫糖尿病（maple syrup urine disease，MSUD）的患病率，分析相关基因突变特点。方法： 采用串联质谱技术对石家庄地区2014年1月—2021年12月出生的185 683例新生儿进行MSUD筛查，对筛查阳性患儿进行BCKDHA、BCKDHB、DBT基因突变检测。结果： 确诊2例MSUD患儿，患病率为1∶92 842。2例患儿的初筛及例2复查串联质谱血液亮氨酸、缬氨酸水平升高，例1因召回复查前已夭折，无复查串联质谱结果。确诊2例MSUD患儿均为经典型，分别检测到BCKDHB和DBT基因复合杂合突变，基因突变分析发现了4种突变位点：c.331C>T、c.289G>T、c.75_76delAT及c.1359_1360delAG；其中c.289G>T和c.1359_1360delAG为未报道基因突变，未检测到BCKDHA基因突变位点。例1于生后10 d夭折；例2于生后8 d开始出现症状，及时干预治疗后好转。结论： 串联质谱技术应用于新生儿疾病筛查可及早发现MSUD患儿；相关基因检测可明确遗传学病因，为遗传咨询提供依据；石家庄地区MSUD的患病率为1∶92 842；发现的4种基因突变位点中2种为未报道基因突变，丰富了基因突变谱。

关键词: 枫糖尿病, 氨基酸代谢障碍, 先天性, 新生儿筛查, 串联质谱法, DNA突变分析

Abstract:

Objective： To explore the prevalence rate of maple syrup urine disease (MSUD) in newborns, and to analyze the mutation characteristics of related genes in Shijiazhuang area, China. Methods： A total of 185 683 newborns born from January 2014 to December 2021 in Shijiazhuang area were screened for MSUD by tandem mass spectrometry, and BCKDHA, BCKDHB and DBT mutations were detected in newborns with positive screening. Results： Two children with MSUD were confirmed, with a prevalence rate of 1∶92 842. In two confirmed cases, the levels of blood leucine (Leu) and valine (Val) were elevated during initial screening. The levels of case 2 were also elevated in the review of tandem mass spectrometry, while case 1 died prematurely before being recalled for review and was not performed the review of tandem mass spectrometry. Two children with MSUD were all classic with gene complex heterozygous mutations of BCKDHB and DBT, and four mutations were found including c.331C>T, c.289G>T, c.75_76delAT and c.1359_1360delAG. Two unreported gene mutations were found, c.289G>T and c.1359_1360delAG. No mutations in the BCKDHA gene were detected. Case 1 died prematurely 10 days after birth. Case 2 began to develop symptoms 8 days after birth, and improved after timely intervention and treatment. Conclusions： Tandem mass spectrometry technology can be applied in newborn disease screening to detect early newborns with MSUD. The genetic testing can identify genetic causes and provide a basis for genetic counseling. The prevalence rate of MSUD in Shijiazhuang area was 1∶92 842. Four gene mutations, two of which were unreported gene mutations, were found in this study, enriching the gene mutation spectrum of MSUD.

Key words: Maple syrup urine disease, Amino acid metabolism, inborn errors, Neonatal screening, Tandem mass spectrometry, DNA mutational analysis

贾立云, 弓苗, 杨会欣, 王熙, 封纪珍. 石家庄地区枫糖尿病患儿串联质谱筛查及BCKDHA、BCKDHB、DBT基因突变分析[J]. 国际生殖健康/计划生育杂志, 2023, 42(3): 203-205.

JIA Li-yun, GONG Miao, YANG Hui-xin, WANG Xi, FENG Ji-zhen. Tandem Mass Spectrometry Screening and BCKDHA, BCKDHB, and DBT Genetic Mutation Analysis in Children with Maple Syrup Urine Disease in Shijiazhuang Area, China[J]. Journal of International Reproductive Health/Family Planning, 2023, 42(3): 203-205.

图/表 2

参考文献 20

[1]	汤欣欣, 郑芹, 刘双, 等. 枫糖尿症1例临床特点及相关基因突变特点分析[J]. 中华实用儿科临床杂志, 2020, 35(20):1586-1588. doi: 10.3760/cma.j.cn101070-20190807-00719. doi: 10.3760/cma.j.cn101070-20190807-00719
[2]	李溪远, 丁圆, 刘玉鹏, 等. 枫糖尿症患儿13例临床、生化及基因研究[J]. 中华实用儿科临床杂志, 2016, 31(8):569-572. doi: 10.3760/cma.j.issn.2095-428X.2016.08.003. doi: 10.3760/cma.j.issn.2095-428X.2016.08.003
[3]	陈正, 罗芳, 吴秀静, 等. 新生儿枫糖尿症二例报告并文献复习[J]. 中华儿科杂志, 2010, 48(9):680-684. doi: 10.3760/cma.j.issn.0578-1310.2010.09.011. doi: 10.3760/cma.j.issn.0578-1310.2010.09.011
[4]	杨彩飞, 陈涛, 雷小光, 等. 枫糖尿病的研究进展[J]. 中华医学遗传学杂志, 2019, 36(7):737-741. doi: 10.3760/cma.j.issn.1003-9406.2019.07.021. doi: 10.3760/cma.j.issn.1003-9406.2019.07.021
[5]	李婕, 梁雁, 罗小平. 枫糖尿症诊治进展[J]. 临床儿科杂志, 2013(7):683-686. doi: 10.3969/j.issn.1000-3606.2013.07.022. doi: 10.3969/j.issn.1000-3606.2013.07.022
[6]	王秋菊, 沈亦平, 陈少科, 等. 遗传变异分类标准与指南[J]. 中国科学(生命科学), 2017, 47(6):668-688. doi: 10.1360/N052017-00099. doi: 10.1360/N052017-00099
[7]	宋东坡, 陈晓英, 吕亚囡, 等. 枫糖尿症患儿基因突变5例分析[J]. 山东医药, 2019, 59(13):58-61. doi: 10.3969/j.issn.1002-266X.2019.13.016. doi: 10.3969/j.issn.1002-266X.2019.13.016
[8]	郭艺, 蒋莉, 孔粼. 经典型枫糖尿症1例随访18个月报告[J]. 临床儿科杂志, 2013(10):968-971. doi: 10.3969/j.issn.1000-3606.2013.10.018. doi: 10.3969/j.issn.1000-3606.2013.10.018
[9]	李晓梅. 枫糖尿症患儿临床特征、基因突变分析及诱导型多能干细胞系的建立[D]. 济南: 山东大学, 2020.
[10]	张璐, 陈晓飞, 朱淑贞. 新生儿枫糖尿病1例的早期护理[J]. 护理与康复, 2016, 15(2):188-190. doi: 10.3969/j.issn.1671-9875.2016.02.034. doi: 10.3969/j.issn.1671-9875.2016.02.034
[11]	黄新文, 张玉, 洪芳, 等. 浙江省新生儿氨基酸代谢疾病筛查及随访分析[J]. 浙江大学学报(医学版), 2017, 46(3):233-239. doi: 10.3785/j.issn.1008-9292.2017.06.02. doi: 10.3785/j.issn.1008-9292.2017.06.02
[12]	马胜举, 赵德华, 马坤, 等. 河南省2013—2019年新生儿遗传代谢病筛查回顾性分析[J]. 检验医学与临床, 2020, 17(14):1965-1968. doi: 10.3969/j.issn.1672-9455.2020.14.006. doi: 10.3969/j.issn.1672-9455.2020.14.006
[13]	鄢慧明, 贾政军, 刘静, 等. 湖南省565182例串联质谱新生儿疾病筛查分析[J]. 中华实用儿科临床杂志, 2019, 34(20):1541-1545. doi: 10.3760/cma.j.issn.2095-428X.2019.20.006. doi: 10.3760/cma.j.issn.2095-428X.2019.20.006
[14]	唐诚芳, 谭敏沂, 谢婷, 等. 广州地区新生儿遗传代谢病串联质谱法筛查结果及筛查性能评估[J]. 浙江大学学报(医学版), 2021, 50(4):463-471. doi: 10.3724/zdxbyxb-2021-0260. doi: 10.3724/zdxbyxb-2021-0260
[15]	Chuang JL, Fisher CR, Cox RP, et al. Molecular basis of maple syrup urine disease: novel mutations at the E1 alpha locus that impair E1(alpha 2 beta 2) assembly or decrease steady-state E1 alpha mRNA levels of branched-chain alpha-keto acid dehydrogenase complex[J]. Am J Hum Genet, 1994, 55(2):297-304. pmid: 8037208
[16]	Wang YP, Qi ML, Li TT, et al. Two novel mutations in the BCKDHB gene(R170H, Q346R) cause the classic form of maple syrup urine disease (MSUD)[J]. Gene, 2012, 498(1):112-115. doi: 10.1016/j.gene.2012.01.082. doi: 10.1016/j.gene.2012.01.082 URL
[17]	Tammachote R, Tongkobpetch S, Desudchit T, et al. Prenatal diagnosis of a novel mutation, c.529C>T(p.Q177X), in the BCKDHA gene in a family with maple syrup urine disease[J]. J Inherit Metab Dis, 2009, 32(Suppl 1):S33-S36. doi: 10.1007/s10545-009-1022-2. doi: 10.1007/s10545-009-1022-2
[18]	Alijanpour M, Jazayeri O, Soleimani Amiri S, et al. Pathogenic Homozygous Mutations in the DBT Gene(c.1174A>C) Result in Maple Syrup Urine Disease in a rs12021720 Carrier[J]. Lab Med, 2022, 53(6):596-601. doi: 10.1093/labmed/lmac034. doi: 10.1093/labmed/lmac034 pmid: 35657820
[19]	Ali EZ, Ngu LH. Fourteen new mutations of BCKDHA, BCKDHB and DBT genes associated with maple syrup urine disease (MSUD) in Malaysian population[J]. Mol Genet Metab Rep, 2018, 17:22-30. doi: 10.1016/j.ymgmr.2018.08.006. doi: 10.1016/j.ymgmr.2018.08.006
[20]	Imtiaz F, Al-Mostafa A, Allam R, et al. Twenty novel mutations in BCKDHA, BCKDHB and DBT genes in a cohort of 52 Saudi Arabian patients with maple syrup urine disease[J]. Mol Genet Metab Rep, 2017, 11:17-23. doi: 10.1016/j.ymgmr.2017.03.006. doi: 10.1016/j.ymgmr.2017.03.006 pmid: 28417071

编号	氨基酸	初筛浓度（μmol/L）	复查浓度（μmol/L）
例1	Leu	1 254.38	-
	Val	627.90	-
例2	Leu	1 088.31	4 182.33
	Val	602.19	998.63

编号	氨基酸	初筛浓度（μmol/L）	复查浓度（μmol/L）
例1	Leu	1 254.38	-
	Val	627.90	-
例2	Leu	1 088.31	4 182.33
	Val	602.19	998.63

编号	基因	基因亚区	基因突变位点	氨基酸改变	纯合/杂合	来源	相关疾病/遗传方式
例1	BCKDHB	EX3	c.331C>T	p.Arg111*	杂合	母亲	MSUDⅠb型/AR
	BCKDHB	EX3	c.289G>T^△	p.Asp97Tyr	杂合	父亲	MSUDⅠb型/AR
例2	DBT	EX2	c.75_76delAT	p.Cys26Trpfs*2	杂合	父亲	MSUDⅡ型/AR
	DBT	EX11	c.1359_1360delAG^△	p.Arg453Serfs*3	杂合	母亲	MSUDⅡ型/AR

编号	基因	基因亚区	基因突变位点	氨基酸改变	纯合/杂合	来源	相关疾病/遗传方式
例1	BCKDHB	EX3	c.331C>T	p.Arg111*	杂合	母亲	MSUDⅠb型/AR
	BCKDHB	EX3	c.289G>T^△	p.Asp97Tyr	杂合	父亲	MSUDⅠb型/AR
例2	DBT	EX2	c.75_76delAT	p.Cys26Trpfs*2	杂合	父亲	MSUDⅡ型/AR
	DBT	EX11	c.1359_1360delAG^△	p.Arg453Serfs*3	杂合	母亲	MSUDⅡ型/AR