新生儿筛查疑诊3-甲基巴豆酰辅酶A羧化酶缺乏症患儿的随访及基因分析
신생인사사의진3-갑기파두선보매A최화매결핍증환인적수방급기인분석
Follow up and gene mutation analysis in cases suspected as 3-methylcrotonyl-coenzyme A carboxylase deficiency by neonatal screening
  • 万方数据
    中华儿科杂志 중화인과잡지
    Chinese Journal of Pediatrics
    2014年 6期 409-414 ,共6页

    叶军%宫丽霏%韩连书%邱文娟%张惠文%高晓岚%金晶%许浩%顾学范

    협군%궁려비%한련서%구문연%장혜문%고효람%금정%허호%고학범

    基因%突变%串联质谱法%新生儿筛查 基因%突變%串聯質譜法%新生兒篩查
    기인%돌변%천련질보법%신생인사사
    Gene%Mutation%Tandem mass spectrometry%Neonatal Screening
    目的 了解新生儿筛查疑诊3-甲基巴豆酰辅酶A羧化酶缺乏症(MCCD)患儿的临床转归;了解中国3-甲基巴豆酰辅酶A羧化酶(MCC)基因突变谱及热点突变.方法 2011年9月至2013年3月,将在上海交通大学医学院附属新华医院接受新生儿筛查,并疑诊为MCCD的42例患儿(男33例,女9例)纳入本研究,根据新生儿筛查召回时血C5-OH浓度、母亲血C5-OH浓度及尿3-MCG、3-HIVA,将42例患儿分为三组:母源性MCCD组5例、良性MCCD组6例、疑似MCCD组31例.对所有患儿的生长、智能发育等随访资料进行回顾分析;对部分患儿进行MCC基因突变检测,分析基因新变异对蛋白结构及功能的影响.结果 (1)42例患儿末次随访的中位年龄29个月,最长随访至9岁.所有患儿临床无症状,生长及智能发育正常.(2)29例患儿(母源性MCCD组4例、良性MCCD组4例,疑似MCCD组21例)接受基因检测,母源性MCCD及良性MCCD基因诊断符合临床诊断;疑似MCCD 21例中仅11例检出1个MCCC1突变,余10例未检出突变.检出的14种突变中MCCC1突变占86％ (12/14),其中c.ins1680A突变占40％,发现9种新突变:211AG＞ CC/p.Q74P、c.295G＞ A/p.G99S、c.764A＞ C/p.H255P、c.964G＞ A/p.E322K、c.1331G＞ A/p.R444H、c.1124delT、c.39_58del20、c.1518delG、c.639 +2T＞ A;5种新错义突变位点多为高度保守氨基酸,经分析这些突变改变了蛋白二级结构,预测对蛋白功能造成影响.(3)母源性MCCD患儿筛查、召回及末次随访的血C5-OH浓度分别为3.50(1.63 ～ 11.43)、1.84(1.00 ～9.30)、0.27(0.26 ～5.81) μmol/L,呈下降趋势;良性MCCD组患儿筛查、召回及末次随访的血C5-OH浓度分别为8.20 (3.60 ～ 9.60)、9.67(3.88 ～20.15)、23.0(5.87 ～49.10) μmol/L,呈上升趋势;疑似MCCD患儿筛查及随访血C5-OH浓度均＜2 μmol/L.良性MCCD组4例患儿召回时尿特异性指标3-MCG均增高.结论 疑诊MCCD患儿最长随访至9岁,临床均无症状、体格及智能发育正常;中国患儿MCCC1突变多见,发现9种新突变,c.ins1680A可能是热点突变.
    목적 료해신생인사사의진3-갑기파두선보매A최화매결핍증(MCCD)환인적림상전귀;료해중국3-갑기파두선보매A최화매(MCC)기인돌변보급열점돌변.방법 2011년9월지2013년3월,장재상해교통대학의학원부속신화의원접수신생인사사,병의진위MCCD적42례환인(남33례,녀9례)납입본연구,근거신생인사사소회시혈C5-OH농도、모친혈C5-OH농도급뇨3-MCG、3-HIVA,장42례환인분위삼조:모원성MCCD조5례、량성MCCD조6례、의사MCCD조31례.대소유환인적생장、지능발육등수방자료진행회고분석;대부분환인진행MCC기인돌변검측,분석기인신변이대단백결구급공능적영향.결과 (1)42례환인말차수방적중위년령29개월,최장수방지9세.소유환인림상무증상,생장급지능발육정상.(2)29례환인(모원성MCCD조4례、량성MCCD조4례,의사MCCD조21례)접수기인검측,모원성MCCD급량성MCCD기인진단부합림상진단;의사MCCD 21례중부11례검출1개MCCC1돌변,여10례미검출돌변.검출적14충돌변중MCCC1돌변점86％ (12/14),기중c.ins1680A돌변점40％,발현9충신돌변:211AG＞ CC/p.Q74P、c.295G＞ A/p.G99S、c.764A＞ C/p.H255P、c.964G＞ A/p.E322K、c.1331G＞ A/p.R444H、c.1124delT、c.39_58del20、c.1518delG、c.639 +2T＞ A;5충신착의돌변위점다위고도보수안기산,경분석저사돌변개변료단백이급결구,예측대단백공능조성영향.(3)모원성MCCD환인사사、소회급말차수방적혈C5-OH농도분별위3.50(1.63 ～ 11.43)、1.84(1.00 ～9.30)、0.27(0.26 ～5.81) μmol/L,정하강추세;량성MCCD조환인사사、소회급말차수방적혈C5-OH농도분별위8.20 (3.60 ～ 9.60)、9.67(3.88 ～20.15)、23.0(5.87 ～49.10) μmol/L,정상승추세;의사MCCD환인사사급수방혈C5-OH농도균＜2 μmol/L.량성MCCD조4례환인소회시뇨특이성지표3-MCG균증고.결론 의진MCCD환인최장수방지9세,림상균무증상、체격급지능발육정상;중국환인MCCC1돌변다견,발현9충신돌변,c.ins1680A가능시열점돌변.
    Objective 3-Methylcrotonyl-coenzyme A carboxylase deficiency (MCCD) is an autosomal recessive inborn error of leucine catabolism.The cases suspected as MCCD detected by neonatal screening are not rare.The aim of the study was to investigate the clinical outcomes in cases suspected as MCCD by neonatal screening.The second aim was to investigate the mutation spectrum of MCC gene in Chinese population and hotspot mutation.Method Forty-two cases (male 33,female 9),who had higher blood 3-hydroxy-isovalerylcarnitine (C5-OH) levels(cut-off ＜0.6 μmol/L) detected by neonatal screening using MS/MS,were recruited to this study during Sept.2011 to Mar.2013.The C5-OH concentrations were [0.84(0.61-20.15) μmol/L] in 42 cases at the screening recall.Five cases were firstly diagnosed as maternal MCCD,6 cases as benign MCCD and 31 cases were suspected as MCCD.To follow up the height,weight,mental development,blood C5-OH concentrations and urinary 3-methylcrotonyl-glycine (3-MCG) and 3-hydroxy isovalerate (3-HIVA) in order to investigate the clinical outcome.The MCCC1 and MCCC2 gene mutation were analyzed for some cases.The novel gene variants were evaluated,and the influence of novel missense variants on the protein structure and function were predicted by PolyPhen-2,SIFT,UniProt and PDB software.Result (1) Forty-two cases had no symptoms,their physical and mental development were normal in the last visit at the median ages of 29 months,the oldest age of follow up was nearly 9 years.(2) Gene mutation analysis was performed for 29 cases with informed consent signed by parents.Fourteen different mutations were identified in 19 cases.The mutations in MCCC1 gene accounted for 86％,the most common mutation was c.ins1680A,(accounted for 40％).Nine kinds of novel variant were detected including 211AG ＞ CC/p.Q74P,c.295G ＞ A/p.G99S,c.764A ＞ C/p.H255P,c.964G ＞ A/p.E322K,c.1331G ＞ A/p.R444H,c.1124delT,c.39_58del20,c.1518delG,c.639 + 2T ＞ A.Other 3 kinds of mutation in MCCC1 gene and 2 kinds of mutation in MCCC2 gene have been reported previously;the amino acid of mutant positions of five kinds of novel missense variant are almost highly conserved.These missense variants were predicted to cause change of human MCC protein side chain structure by changing hydrogen bonding,size of amino acid residue and electric charge,and predicted to damage the protein function possibly according to PolyPhen-2 and PDB analysis.So these novel variants may be disease-causing mutations.No mutation were detected in 10 cases.(3) Blood concentrations of C5-OH when screening,recall and end of follow-up in maternal MCCD was 3.50 (1.63-11.43),1.84 (1.00-9.30),0.27 (0.26-5.81) μmol/L.There was a significant downward trend.In contrast,benign MCCD group was 8.20 (3.60-9.60),9.67 (3.88-20.15),23.0(5.87-49.10) μmol/L.It showed a rising trend.Children's urinary 3-MCG of benign MCCD group was found abnormally elevated in 4 cases (100％) when they were recalled.Conclusion A certain number of cases with MCCD or suspected as MCCD in this study had no symptoms and normal physical and mental development after follow-up to oldest age of nearly 9 years.The mutation in MCCC1 gene is common,nine novel mutations were found,c.ins1680A may be a hotspot mutation in Chinese population.The urinary GC/MS analysis and blood MS/MS analysis for mother should be routinely performed for all cases with high blood C5-OH level detected by neonatal screening.
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