新生儿高脯氨酸血症串联质谱筛查结果及基因突变分析

doi:10.12280/gjszjk.20210433

摘要/Abstract

摘要：

目的： 回顾性分析石家庄市新生儿高脯氨酸血症筛查情况,了解其患病率及脯氨酸脱氢酶（proline dehydrogenase,PRODH）突变情况。方法： 采用串联质谱技术-非衍生法检测滤纸干血斑中脯氨酸浓度,对石家庄市2014年1月—2020年12月出生的活产新生儿进行筛查,进一步采用基因测序技术对筛查阳性患儿进行PRODH基因突变检测,Sanger测序验证。结果： 172 895例新生儿中筛查阳性患儿10例,基因确诊5例,患病率为1/34 579。5例患儿中PRODH基因复合杂合突变4例;基因整体杂合缺失1例。基因突变分析发现点突变4种,其中文献已报道2种,分别为c.1322T>C和c.1363G>T;文献尚未报道2种,分别为c.273+1G>C和c.334delA。随访结果发现,除1例患儿语言发育稍落后,其余均正常。结论： 石家庄市新生儿高脯氨酸血症患病率为1/34 579;检测到基因突变确诊5例患儿,发现2种未报道基因突变,丰富了基因数据库。

关键词: 新生儿筛查, 串联质谱法, 高脯氨酸血症, 基因, 突变

Abstract:

Objective: To retrospectively analyze the screening status of neonatal hyperprolinemia in Shijiazhuang City, so as to understand its prevalence and proline dehydrogenase (PRODH) mutations. Methods: Tandem mass spectrometry technology-non-derivatization method was used to detect proline levels in dried blood spots on filter paper. Live birth newborns born between January 2014 and December 2020 in Shijiazhuang City were screened. Genetic sequencing technology was further used to detect the mutations of the PRODH gene in children with positive screening. Sanger sequencing was used for verification. Results: Ten of the 172 895 newborns were screened suspected positive, 5 cases were genetically confirmed, with the incidence rate 1/34 579. Among the 5 children, 4 cases were compound heterozygous mutations in PRODH gene, 1 case was loss of heterozygous gene overall. Gene mutation analysis revealed four kinds of point mutations, in which two mutations, c.1322T>C and c.1363G>T, have been reported. Two mutations, c.273+1G>C and c.334delA, were not yet reported. Conclusions: The prevalence of neonatal hyperprolinemia in Shijiazhuang City was 1/34 579. Five cases of children were diagnosed with gene mutations, and two unreported gene mutations were found, which enriched the gene database.

Key words: Neonatal screening, Tandem mass spectrometry, Hyperprolinemia, Genes, Mutation

封露露, 贾立云, 弓苗, 马翠霞, 封纪珍. 新生儿高脯氨酸血症串联质谱筛查结果及基因突变分析[J]. 国际生殖健康/计划生育, 2022, 41(1): 18-21.

FENG Lu-lu, JIA Li-yun, GONG Miao, MA Cui-xia, FENG Ji-zhen. Tandem Mass Spectrometry Screening and Gene Mutation Analysis of Neonatal Hyperprolinemia[J]. Journal of International Reproductive Health/Family Planning, 2022, 41(1): 18-21.

图/表 4

参考文献 22

[1]	王彦云, 孙云, 程威, 等. 高脯氨酸血症临床特征分析[J]. 临床儿科杂志, 2020, 38(1):40-43. doi: 10.3969/j.issn.1000-3606.2020. 01.010. doi: 10.3969/j.issn.1000-3606.2020. 01.010
[2]	廉文君, 郑宏, 陆相朋, 等. 高脯氨酸血症1例报告[J]. 临床儿科杂志, 2019, 37(11):862-865. doi: 10.3969/j.issn.1000-3606.2019.11.016. doi: 10.3969/j.issn.1000-3606.2019.11.016
[3]	Kaur R, Paria P, Saini AG, et al. Metabolic epilepsy in hyperprolinemia type II due to a novel nonsense ALDH4A1 gene variant[J]. Metab Brain Dis, 2021, 36(6):1413-1417. doi: 10.1007/s11011-021-00757-w. doi: 10.1007/s11011-021-00757-w URL
[4]	Ersoy M, Ylmaz S, Ceylaner S. Antioxidant Therapy in a Patient with Hyperprolinemia Type 1 Presenting with Mild Neuromotor Retardation and Speech Disturbance[J]. Indian J Pediatr, 2021, 88(6):601. doi: 10.1007/s12098-021-03744-2. doi: 10.1007/s12098-021-03744-2 URL
[5]	Liu LK, Becker DF, Tanner JJ. Structure, function, and mechanism of proline utilization A(PutA)[J]. Arch Biochem Biophys, 2017, 632:142-157. doi: 10.1016/j.abb.2017.07.005. doi: S0003-9861(17)30363-6 pmid: 28712849
[6]	沈玉燕, 黎剑, 肖刚. 怀化地区新生儿氨基酸代谢病筛查及随访分析[J]. 中国优生与遗传杂志, 2018, 26(10):80-82.
[7]	陆海燕, 郑建丽, 姚艳, 等. 盐城地区新生儿遗传代谢病串联质谱筛查结果回顾性分析[J]. 中国优生与遗传杂志, 2016, 24(3):83-85.
[8]	孙云, 王彦云, 马定远, 等. 南京地区175 767例串联质谱技术新生儿筛查结果分析[J]. 中华围产医学杂志, 2020, 23(4):224-231. doi: 10.3760/cma.j.cn113903-20190428-00289. doi: 10.3760/cma.j.cn113903-20190428-00289
[9]	鞠彩宁, 蓝信强. 串联质谱技术在威海地区44438例新生儿遗传代谢病筛查中的应用[J]. 医学检验与临床, 2020, 31(9):43-45. doi: 10.3969/j.issn.1673-5013.2020.09.011. doi: 10.3969/j.issn.1673-5013.2020.09.011
[10]	裴薇, 鹿相花, 田维兵, 等. 潍坊地区新生儿遗传代谢病串联质谱筛查结果分析[J]. 中国中西医结合儿科学, 2018, 10(5):452-455. doi: 10.3969/j.issn.1674-3865.2018.05.025. doi: 10.3969/j.issn.1674-3865.2018.05.025
[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]	钟锦平, 彭维林, 傅清流, 等. 福建泉州地区新生儿氨基酸代谢障碍的筛查结果分析[J]. 现代检验医学杂志, 2020, 35(4):41-44,78. doi: 10.3969/j.issn.1671-7414.2020.04.010. doi: 10.3969/j.issn.1671-7414.2020.04.010
[13]	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
[14]	沈玉燕, 黎剑, 肖刚. 高脯氨酸血症Ⅰ型伴运动落后患儿一例[J]. 中国优生与遗传杂志, 2018, 26(4):74,86.
[15]	Namavar Y, Duineveld DJ, Both GIA, et al. Psychiatric phenotypes associated with hyperprolinemia: A systematic review[J]. Am J Med Genet B Neuropsychiatr Genet, 2021, 186(5):289-317. doi: 10.1002/ajmg.b.32869. doi: 10.1002/ajmg.b.32869 URL
[16]	Hama R, Kido J, Sugawara K, et al. Hyperprolinemia type I caused by homozygous p.T466M mutation in PRODH[J]. Hum Genome Var, 2021, 8(1):28. doi: 10.1038/s41439-021-00159-5. doi: 10.1038/s41439-021-00159-5 URL
[17]	Afenjar A, Moutard ML, Doummar D, et al. Early neurological phenotype in 4 children with biallelic PRODH mutations[J]. Brain Dev, 2007, 29(9):547-552. doi: 10.1016/j.braindev.2007.01.008. doi: 10.1016/j.braindev.2007.01.008 URL
[18]	Mitsubuchi H, Nakamura K, Matsumoto S, et al. Inborn Errors of Proline Metabolism[J]. J Nutr, 2008, 138(10):2016-2020. doi: 10.1111/j.1365-277X.2008.00912.x. doi: 10.1111/j.1365-277X.2008.00912.x pmid: 18806117
[19]	Duarte M, Afonso J, Moreira A, et al. Hyperprolinemia as a clue in the diagnosis of a patient with psychiatric manifestations[J]. Brain Dev, 2017, 39(6):539-541. doi: 10.1016/j.braindev.2017.01.008. doi: 10.1016/j.braindev.2017.01.008 URL
[20]	Di Rosa G, Pustorino G, Spano M, et al. Type I hyperprolinemia and proline dehydrogenase (PRODH) mutations in four Italian children with epilepsy and mental retardation[J]. Psychiatr Genet, 2008, 18(1):40-42. doi: 10.1097/YPG.0b013e3282f08a3d. doi: 10.1097/YPG.0b013e3282f08a3d URL
[21]	Jacquet H, Raux G, Thibaut F, et al. PRODH mutations and hyperprolinemia in a subset of schizophrenic patients[J]. Hum Mol Genet, 2002, 11(19):2243-2249. doi: 10.1093/hmg/11.19.2243. doi: 10.1093/hmg/11.19.2243 pmid: 12217952
[22]	Cappelletti P, Tallarita E, Rabattoni V, et al. Proline oxidase controls proline, glutamate, and glutamine cellular concentrations in a U87 glioblastoma cell line[J]. PLoS One, 2018, 13(4):e0196283. doi: 10.1371/journal.pone.0196283.eCollection 2018. doi: 10.1371/journal.pone.0196283.eCollection 2018 URL

病例	性别	初筛	召回复查
1	女	573.16	428.34
2	女	331.82	-
3	男	91.19	349.72
4	男	853.55	400.70
5	女	568.35	766.65
6	女	399.24	518.31
7	女	409.01	737.56
8	女	424.60	638.73
9	男	616.01	509.46
10	男	560.06	720.50

病例	性别	初筛	召回复查
1	女	573.16	428.34
2	女	331.82	-
3	男	91.19	349.72
4	男	853.55	400.70
5	女	568.35	766.65
6	女	399.24	518.31
7	女	409.01	737.56
8	女	424.60	638.73
9	男	616.01	509.46
10	男	560.06	720.50

病例	基因	转录本	Exon/Intron	核苷酸	氨基酸	杂合/纯合	突变来源
1	PRODH	NM_016335	Exon12	c.1322T>C	p.L441P	杂合	父亲
		NM_016335	Intron2	c.273+1G>C^☆	p.?	杂合	母亲
2	PRODH	NM_016335	Exon3	c.334delA^☆	p.T112Pfs*7	杂合	父亲
		NM_016335	Exon12	c.1322T>C	p.L441P	杂合	母亲
3	PRODH	基因整体杂合缺失
4	PRODH	NM_016335	Exon12	c.1322T>C	p.L441P	杂合	母亲
		NM_016335	Exon12	c.1363G>T	p.A455S	杂合	父亲
5	PRODH	NM_016335	Exon12	c.1363G>T	p.A455S	杂合	父亲
		NM_016335	Exon12	c.1322T>C	p.L441P	杂合	母亲

病例	基因	转录本	Exon/Intron	核苷酸	氨基酸	杂合/纯合	突变来源
1	PRODH	NM_016335	Exon12	c.1322T>C	p.L441P	杂合	父亲
		NM_016335	Intron2	c.273+1G>C^☆	p.?	杂合	母亲
2	PRODH	NM_016335	Exon3	c.334delA^☆	p.T112Pfs*7	杂合	父亲
		NM_016335	Exon12	c.1322T>C	p.L441P	杂合	母亲
3	PRODH	基因整体杂合缺失
4	PRODH	NM_016335	Exon12	c.1322T>C	p.L441P	杂合	母亲
		NM_016335	Exon12	c.1363G>T	p.A455S	杂合	父亲
5	PRODH	NM_016335	Exon12	c.1363G>T	p.A455S	杂合	父亲
		NM_016335	Exon12	c.1322T>C	p.L441P	杂合	母亲

病例	随访截止时年龄	饮食治疗	药物治疗	随访截止时脯氨酸浓度（μmol/L）	生长发育情况
1	5岁7个月	6个月	无	805	正常
2	4岁2个月	无	无	426	正常
3	3岁6个月	无	无	457	正常
4	3岁10个月	3个月	无	639	正常
5	1岁5个月	无	无	4 200	语言稍落后