尿素循环障碍患者家系的基因突变分析及产前诊断

doi:10.12280/gjszjk.20200537

摘要/Abstract

摘要：

目的:分析7例尿素循环障碍(urea cycle disorder,UCD)患者家系的基因突变情况,并对2例再生育家庭进行产前诊断。方法:联合应用高通量测序(panel)结合Sanger测序、长距离-聚合酶链反应(LD-PCR)以及多重连接探针扩增(multiplex ligation-dependent probe amplification,MLPA)等基因检测技术,对7例疑似UCD患者家系进行相关致病基因突变分析,并对其中2例高风险家系的胎儿羊水标本进行产前基因诊断。结果:7例疑似UCD患者均得到明确基因诊断:4例患儿为SLC25A13基因突变引起的新生儿肝内胆汁淤积症(neonatal intrahepatic cholestasis caused by citrin deficiency,NICCD),且1例同时患脊髓性肌萎缩症(spinal muscular atrophy,SMA);1例患儿为ASS1基因突变引起的瓜氨酸血症Ⅰ型(citrullinaemia typeⅠ,CTLN1);2例患儿为OTC基因突变引起的鸟氨酸氨甲酰转移酶缺乏症(ornithine transcarbamylase deficiency,OTCD)。2家系再生育时产前诊断结果:1例胎儿为父源SLC25A13致病基因携带者;1例为OTC正常的UCD胎儿。结论:基因诊断有助于可疑UCD患者的明确诊断以及分型,部分疑似UCD胎儿可能OTC正常。对于生育过UCD患儿的家系,再生育时进行产前基因诊断可积极预防出生缺陷。

关键词: 尿素循环障碍,先天性, 瓜氨酸血症, 鸟氨酸氨甲酰转移酶缺乏症, DNA突变分析, 产前诊断, 新生儿肝内胆汁淤积症

Abstract:

Objective: To investigate the gene mutations of seven families suspected with urea cycle disorders (UCD), and to perform prenatal diagnosis for two reproductive families. Methods: Target sequence capture combined with high-throughput next-generation sequencing, and Sanger sequencing/LD-PCR/MLPA (multiplex ligation-dependent probe amplification) technologies were used to investigate the gene mutations for seven families suspected with UCD. After the genotypes of the pedigrees were identified, two amniotic fluid samples from high-risk pregnant women were used for prenatal genetic diagnosis. Results: The diagnosis of UCD was confirmed in seven suspected families. Among of them, four patients were diagnosed as the neonatal intrahepatic cholestasis caused by citrin deficiency (NICCD) and one of them was coexist with spinal muscular atrophy (SMA). One patient was diagnosed with citrullinaemia typeⅠ(CTLN1). And two patients were diagnosed with ornithine transcarbamylase deficiency (OTCD). Prenatal genetic diagnosis of two families show that one fetus had a mutation at a heterozygous mutation of SLC25A13 with paternal; and another one was a proband with normal OTC gene. Conclusions: Genetic diagnostic can make accurate and effective diagnosis of UCD. However, some families with suspected UCD could have normal OTC gene. The prenatal genetic diagnosis combined with OTC gene testing can prevent the rebirth defects in high-risk families.

Key words: Urea cycle disorders,inborn, Citrullinemia, Ornithine carbamoyltransferase deficiency disease, DNA mutational analysis, Prenatal diagnosis, Neonatal intrahepatic cholestasis caused by citrin deficiency

张庆华, 郝胜菊, 王兴, 陈雪, 周秉博, 刘芙蓉, 郑雷, 冯暄, 张钏. 尿素循环障碍患者家系的基因突变分析及产前诊断[J]. 国际生殖健康/计划生育, 2021, 40(3): 185-188.

ZHANG Qing-hua, HAO Sheng-ju, WANG Xing, CHEN Xue, ZHOU Bing-bo, LIU Fu-rong, ZHENG Lei, FENG Xuan, ZHANG Chuan. Gene Mutation Analysis and Prenatal Diagnosis of Family with Urea Cycle Disorder[J]. Journal of International Reproductive Health/Family Planning, 2021, 40(3): 185-188.

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序号	性别	临床表型	基因	基因型(父/母)			疾病名称	遗传模式
序号	性别	临床表型	基因	父源性	母源性	新发性	疾病名称	遗传模式
1	女	颈部柔软,无抵抗,皮肤巩膜黄染,高胆红素血症	SLC25A13	-	IVS16ins3kb	c.1737G>A	NICCD合并SMA	AR
			SMN1	E7-8del	E7-8del	-		AR
2	男	胆汁淤积,黄疸,肉碱代谢异常,瓜氨酸代谢异常	SLC25A13	c.1364G>T	c.1660_1661ins16bp	-	NICCD	AR
3	女	皮肤黄染,高胆红素血症,瓜氨酸指标异常	SLC25A13	c.1638_1660dup23	IVS16ins3kb	-	NICCD	AR
4	男	胆汁淤积性肝病,血液蛋白不足,低纤维蛋白原血症	SLC25A13	c.1638_1660dup23	IVS16ins3kb	-	NICCD	AR
5	男	高氨血症,新生儿惊厥,呼吸暂停	ASS1	c.1088G>A	c.787G>A	-	CTLN1	AR
6	男	新生儿惊厥,代谢性酸中毒,高氨酸血症	OTC	-	-	c.958C>T	OTCD	XL
7	女	高氨血症,尿素循环障碍,呼吸衰竭	OTC	-	Exon7-9 Del	-	OTCD	XL

序号	性别	临床表型	基因	基因型(父/母)			疾病名称	遗传模式
序号	性别	临床表型	基因	父源性	母源性	新发性	疾病名称	遗传模式
1	女	颈部柔软,无抵抗,皮肤巩膜黄染,高胆红素血症	SLC25A13	-	IVS16ins3kb	c.1737G>A	NICCD合并SMA	AR
			SMN1	E7-8del	E7-8del	-		AR
2	男	胆汁淤积,黄疸,肉碱代谢异常,瓜氨酸代谢异常	SLC25A13	c.1364G>T	c.1660_1661ins16bp	-	NICCD	AR
3	女	皮肤黄染,高胆红素血症,瓜氨酸指标异常	SLC25A13	c.1638_1660dup23	IVS16ins3kb	-	NICCD	AR
4	男	胆汁淤积性肝病,血液蛋白不足,低纤维蛋白原血症	SLC25A13	c.1638_1660dup23	IVS16ins3kb	-	NICCD	AR
5	男	高氨血症,新生儿惊厥,呼吸暂停	ASS1	c.1088G>A	c.787G>A	-	CTLN1	AR
6	男	新生儿惊厥,代谢性酸中毒,高氨酸血症	OTC	-	-	c.958C>T	OTCD	XL
7	女	高氨血症,尿素循环障碍,呼吸衰竭	OTC	-	Exon7-9 Del	-	OTCD	XL

基因名称	碱基改变	氨基酸改变	突变类型	频数	HGMD (+/-)	clinvar (+/-)
SLC25A13	c.1737G>A	p.W579X	无义突变	1	+	-
	IVS16ins3kb	-	插入突变	3	+	+
	c.1638_1660dup23	p.A554Gfs*17	移码突变	2	+	+
	c.1364G>T	p.R455L	错义突变	1	+	-
	c.1660_1661ins16bp	p.A554Gfs*17	移码突变	1	-	+
ASS1	c.787G>A	p.V263M	错义突变	1	+	+
	c.1088G>A	p.R363Q	错义突变	1	+	+
OTC	c.958C>T	p.R320X	无义突变	1	+	-
	Exon7-9 Del	-	缺失突变	1	+	+

基因名称	碱基改变	氨基酸改变	突变类型	频数	HGMD (+/-)	clinvar (+/-)
SLC25A13	c.1737G>A	p.W579X	无义突变	1	+	-
	IVS16ins3kb	-	插入突变	3	+	+
	c.1638_1660dup23	p.A554Gfs*17	移码突变	2	+	+
	c.1364G>T	p.R455L	错义突变	1	+	-
	c.1660_1661ins16bp	p.A554Gfs*17	移码突变	1	-	+
ASS1	c.787G>A	p.V263M	错义突变	1	+	+
	c.1088G>A	p.R363Q	错义突变	1	+	+
OTC	c.958C>T	p.R320X	无义突变	1	+	-
	Exon7-9 Del	-	缺失突变	1	+	+