目的 分析4 个甲基丙二酸尿症（MMA）的家系基因突变情况，阐明开展MMA 基因突变分析及产前诊断的意义。方法 对诊断为MMA 的先证者或其父母行相关基因的高通量测序，确定基因突变位点，并采用聚合酶链反应和直接测序法对家系行一代测序验证。家系1、3、4 于先证者母亲再次妊娠11~13 周时超声引导下行绒毛活检，进行早期产前诊断。结果 家系1 先证者父亲检出MUT 基因c.656A > T 杂合突变，先证者母亲检出c.729-730insTT 杂合突变；早期产前诊断发现胎儿为c.656A > T 和c.729-730insTT 双重杂合突变，终止妊娠。家系2 先证者检出MUT 基因c.1106G > A 和c.755-756insA 双重杂合突变，c.1106G > A 来自父亲，c.755-756insA 来自母亲。家系3 先证者检出MMACHC 基因c.217C > T 和c.609G > A 双重杂合突变，c.217C > T 来自父亲，c.609G > A 来自母亲；产前诊断提示胎儿携带c.609G > A 杂合突变，胎儿出生时脐血检测结果与产前诊断一致。家系4 先证者检出MMACHC 基因c.609G > A 和c.567dupT 双重杂合突变，c.609G > A 来自父亲，c.567dupT 来自母亲；产前诊断提示胎儿携带c.567dupT 杂合突变，胎儿顺利出生，脐血检测结果与产前诊断一致。结论 明确基因突变有助于MMA 家系行产前诊断，避免缺陷患儿出生。
Abstract
Objective To study gene mutations in four pedigrees with methymalonic aciduria, as well as the feasibility of prenatal diagnosis of methymalonic aciduria. Methods High-throughput sequencing was performed for related genes in the peripheral blood of children or parents who were diagnosed with methymalonic aciduria to identify the loci with mutations. Then amplification primers were designed for each locus, and PCR and direct sequencing were performed to validate the sequencing in the first generation in the four pedigrees. Whether the mutations were pathogenic were determined with reference to literature review and medical history. In the pedigrees 1, 3, and 4, ultrasound-guided chorionic villi biopsy was performed at weeks 11-13 of pregnancy to perform early prenatal diagnosis. Results In pedigree 1, c.656A > T and c.729-730insTT heterozygous mutations in the MUT gene were detected in the proband’s father and mother, respectively. Early prenatal diagnosis showed c.656A > T and c.729-730insTT double heterozygous mutations in the fetus. The couple decided to terminate pregnancy. In pedigree 2, c.1106G > A and c.755-756insA double heterozygous mutations in the MUT gene were detected in the proband. c.1106G > A came from the father and c.755-756insA came from the mother. In pedigree 3, c.217C > T and c.609G > A double heterozygous mutations in the MMACHC gene were detected in the proband. c.217C > T came from the father and c.609G > A came from the mother. Prenatal diagnosis showed c.609G > A heterozygous mutation in the fetus. The baby was successfully delivered, and the result of umbilical cord blood testing was consistent with the prenatal diagnosis. In pedigree 4, c.609G > A and c.567dupT double heterozygous mutations in the MMACHC gene were detected in the proband. c.609G > A came from the father and c.567dupT came from the mother. Prenatal diagnosis showed c.567dupT heterozygous mutation in the fetus. The baby was successfully delivered, and the result of umbilical cord blood testing was consistent with the prenatal diagnosis.Conclusions Identification of gene mutations helps with prenatal diagnosis in pedigrees with methymalonic aciduria.
关键词
甲基丙二酸尿症 /
基因突变 /
产前诊断 /
家系研究
Key words
Methymalonic aciduria /
Gene mutation /
Prenatal diagnosis /
pedigree study
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参考文献
[1] Oberholzer VG, Levin B, Burgess EA, et al. Methylmalonic aciduria. An inborn error of metabolism leading to chronic metabolic acidosis[J]. Arch Dis Child, 1967, 42(225): 492-504.
[2] Han B, Cao Z, Tian L, et al. Clinical presentation, gene analysis and outcomes in young patients with early-treated combined methylmalonic acidemia and homocysteinemia (cblC type) in Shandong province, China[J]. Brain Dev, 2016, 38(5): 491-497.
[3] Vatanavicharn N, Champattanachai V, Liammongkolkul S, et al. Clinical and molecular findings in Thai patients with isolated methylmalonic acidemia[J]. Mol Genet Metab, 2012, 106(4): 424-429.
[4] Dündar H, Özgül RK, Güzel-Ozantürk A, et al. Microarray based mutational analysis of patients with methylmalonic acidemia: identification of 10 novel mutations[J]. Mol Genet Metab, 2012, 106(4): 419-423.
[5] Wang J, Li E, Wang L, et al. Genetic analysis of four cases of methylmalonic aciduria and homocystinuria, cblC type#[J]. Int J Clin Exp Pathol, 2015, 8(8): 9337-9341.
[6] Weisfeld-Adams JD, Morrissey MA, Kirmse BM, et al. Newborn screening and early biochemical follow-up in combined methylmalonic aciduria and homocystinuria, cblC type, and utility of methionine as a secondary screening analyte[J]. Mol Genet Metab, 2010, 99(2): 116-123.
[7] Lempp TJ, Suormala T, Siegenthaler R, et al. Mutation and biochemical analysis of 19 probands with mut0 and 13 with mut-methylmalonic aciduria: identification of seven novel mutations[J]. Mol Genet Metab, 2007, 90(3): 284-290.
[8] Shi C, Shang D, Sun S, et al. MMACHC gene mutation in familial hypogonadism with neurological symptoms[J]. Gene, 2015, 574(2): 380-384.
[9] Nogueira C, Aiello C, Cerone R, et al. Spectrum of MMACHC mutations in Italian and Portuguese patients with combined methylmalonic aciduria and homocystinuria, cblC type[J]. Mol Genet Metab, 2008, 93(4): 475-480.
[10] Wang X, Sun W, Yang Y, et al. A clinical and gene analysis of late-onset combined methylmalonic aciduria and homocystinuria, cblC type, in China[J]. J Neurol Sci, 2012, 318(1-2): 155-159.
[11] Cosson MA, Benoist JF, Touati G, et al. Long-term outcome in methylmalonic aciduria: a series of 30 French patients[J]. Mol Genet Metab, 2009, 97(3): 172-178.
[12] Morel CF, Lerner-Ellis JP, Rosenblatt DS. Combined methylmalonic aciduria and homocystinuria (cblC): phenotypegenotype correlations and ethnic-specific observations[J]. Mol Genet Metab, 2006, 88(4): 315-321.
[13] Ghoraba DA, Mohammed MM, Zaki OK. Mutation analysis of methylmalonyl CoA mutase gene exon 2 in Egyptian families: Identification of 25 novel allelic variants[J]. Meta Gene, 2015, 3: 71-88.
[14] Acquaviva C, Benoist JF, Callebaut I, et al. N219Y, a new frequent mutation among mut(degree) forms of methylmalonic acidemia in Caucasian patients[J]. Eur J Hum Genet, 2001, 9(8): 577-582.
[15] Lerner-Ellis JP, Tirone JC, Pawelek PD, et al. Identification of the gene responsible for methylmalonic aciduria and homocystinuria, cblC type[J]. Nat Genet, 2006, 38(1): 93-100.
[16] Chang JT, Chen YY, Liu TT, et al. Combined methylmalonic aciduria and homocystinuria cblC type of a Taiwanese infant with c.609G > A and C.567dupT mutations in the MMACHC gene[J]. Pediatr Neonatol, 2011, 52(4): 223-226.
[17] 王斐, 韩连书, 叶军, 等. 甲基丙二酸血症患儿MUT 基因突 变分析[J]. 中华医学遗传学杂志, 2009, 26(5): 485-489.
[18] 王斐, 韩连书, 胡宇慧, 等. 甲基丙二酸血症伴同型半胱氨 酸血症患儿基因突变分析[J]. 中华儿科杂志, 2009, 47(3): 189-193.
[19] 韩凤, 韩连书, 王瑜, 等. 质谱技术检测羊水中代谢物产前 诊断甲基丙二酸血症[J]. 中华围产医学杂志, 2015, 18(3): 182-187.
[20] 刘宁, 史惠蓉, 孔祥东, 等. MUT 基因突变致单纯性甲基丙 二酸尿症家系的突变分析及产前诊断[J]. 中华医学遗传学杂 志, 2013, 30(6): 747-749.
[21] Zong Y, Liu N, Zhao Z, et al. Prenatal diagnosis using genetic sequencing and identification of a novel mutation in MMACHC[J]. BMC Med Genet, 2015, 16: 48.
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