单核苷酸多态性微阵列技术在高龄孕妇产前诊断中的应用

苏景玉; 欧阳鲁平; 欧珊; 刘天盛; 李薇; 费冬梅; 黄红倩; 耿国兴; 罗静思

doi:10.16766/j.cnki.issn.1674-4152.002988

单核苷酸多态性微阵列技术在高龄孕妇产前诊断中的应用

doi: 10.16766/j.cnki.issn.1674-4152.002988

广西壮族自治区妇幼保健院遗传代谢中心实验室，广西壮族自治区出生缺陷预防控制研究所, 广西南宁 530002

基金项目:

广西自然科学基金项目 2020JJB140047

广西卫健委自筹课题 Z20210440

广西卫健委自筹课题 Z20210066

广西卫健委自筹课题 Z20200699

详细信息

通讯作者:
罗静思，E-mail：luojingsi0815@126.com

中图分类号: R714.5 R446
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出版历程
- 收稿日期: 2022-01-03

Application of single nucleotide polymorphic microarray technique in prenatal diagnosis of advanced maternal age

Laboratory of Genetic and Metabolism Center, Maternal & Child Health Hospital of Guangxi Zhuang Autonomous Region, Guangxi Institute of Birth Defects Prevention and Control, Nanning, Guangxi 530002, China

摘要

摘要: 目的探讨染色体核型与单核苷酸多态性微阵列技术(SNP array)在高龄妊娠孕妇胎儿的产前诊断中联合应用的临床价值及染色体变异位点与相应临床表型的关联性。方法统计2016年1月—2020年12月来广西壮族自治区妇幼保健院遗传门诊就诊的2 642例因高龄妊娠(≥35岁)而行羊膜腔穿刺抽取羊水的孕妇, 采用常规的传代细胞培养, 行染色体核型分析, G显带; 同时并行SNP array技术检测染色体拷贝数变异(copy number variations, CNVs)。结果共2 642例高龄孕妇羊水样本进行核型分析，检出44例非整倍体染色体异常，其中21-三体综合征最多(65.9%, 29/44)，其次为18-三体综合征(9.1%, 4/44)、13-三体综合征(4.5%, 2/44)、性染色体非整倍体嵌合(20.5%, 9/44)。SNP array技术还检出20例染色体微缺失或微重复异常，以及104例临床意义未明病例。检测率方面比较，SNP array检测明显高于核型分析技术(P < 0.05)。核型分析在年龄段中的比较：35~37岁年龄段异常检出率与38~40岁年龄段；以及38~40岁年龄段与41~43岁年龄段差异均无统计学意义(均P>0.05)，41~43岁年龄段异常检出率与≥44岁年龄段差异有统计学意义(P < 0.05)。SNP array技术在高龄妊娠这4个年龄段检出率差异均无统计学意义(P>0.05)。结论高龄妊娠与胎儿染色体异常相关，以染色体非整倍体为主，但也存在CNVs、易位等染色体异常。作为高龄妊娠的胎儿，同时进行染色体核型分析和SNP array检测，可以为高龄妊娠胎儿的遗传咨询和预后评估提供更为精准的信息和依据。
- 高龄孕妇 /
- 单核苷酸多态性微阵列芯片 /
- 染色体核型分析 /
- 产前诊断
Abstract: Objective To investigate the clinical value of the combination of chromosome karyotype and single nucleodide polymorphic microarray (SNP array) in the antenatal diagnosis of fetuses with advanced pregnancy and the correlation between chromosome loci and corresponding clinical phenotypes. Methods From January 2016 to December 2020, 2 642 pregnant women who underwent amniocentesis to extract amniotic fluid due to advanced pregnancy (≥35 years old) were enrolled in the genetic outpatient clinic of Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region. Conventional cell culture was used for chromosome karyotype analysis and G-banding. At the same time, copy number variations (CNVs) were detected by parallel SNP array. Results Karyotype analysis of 2 642 amniotic fluid samples from advanced pregnant women revealed 44 aneuploidy chromosome abnormalities, among which trisomy 21-syndrome was the most common (65.9%, 29/44), followed by trisomy 18-syndrome (9.1%, 4/44), trisomy 13-syndrome (4.5%, 2/44), and sex chromosome aneuploidy chimerism (20.5%, 9/44). The SNP array also detected 20 cases of chromosomal microdeletion or microduplication abnormalities and 104 cases of unknown clinical significance. The detection rate of SNP array was significantly higher than that of karyotype analysis (P < 0.05). Comparison of karyotype analysis in age groups: the difference between the karyotype analysis in the age groups was not statistically significant (P>0.05) between the 35-37 years old and 38-40 years old age group; and between the 38-40 years old and 41-43 years old age group(P>0.05), while the difference between 41-43 years old and ≥44 years old age group was statistically significant (P < 0.05). There was no significant difference in SNP array detection rates in the four ages of advanced pregnancy (P>0.05). Conclusion The fetus of advanced maternal age is associated with chromosomal abnormalities, mainly chromosomal aneuploidy, but also CNVs, translocation and other chromosomal abnormalities. Simultaneous chromosome karyotype analysis and SNP array detection can provide more accurate information and basis for genetic counselling and prognostic assessment of fetus with advanced pregnancy.
- Advanced maternal age /
- Single nucleotide polymorphism microarray /
- Karyotype analysis /
- Prenatal diagnosis
利益冲突 无

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表 1 各年龄段染色体核型分析检出的非整倍体情况(例)

Table 1. Aneuploidy detected by karyotype analysis at different ages(n)

年龄(岁)	总非整倍体	21-三体	18-三体	13-三体	性染色体非整倍体
35~37	11	5	1	2	3
38~40	12	9	1	0	2
41~43	14	9	1	0	4
≥44	7	6	1	0	0
合计	44	29	4	2	9

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表 2 SNP array额外检出的20例微缺失、微重复病例情况

Table 2. Additional 20 cases of microdeletion and microduplication detected by SNP array

病例	年龄(岁)	检出拷贝数变异	数据库查询结果	妊娠结局
1	35	arr[hg19]1q21.1q21.2(146476526-147825227)x3	致病性CNVs，涉及1q21.1重复综合征，包含关键基因PRKAB2和GJA5。	终止妊娠
2	37	arr[hg19]22q11.21(18844632-21462353)x3	致病性CNVs, 涉及22q11.2重复综合征，临床表现精神运动发育迟缓，异常面容等。	终止妊娠
3	37	arr[hg19]16p13.11(15052746-16303388)x1	致病性CNVs，临床表现全面性发育迟缓，主动脉缩窄，神经系统异常，马蹄足内翻，面部畸形等。	终止妊娠
4	37	arr [hg19]Xp22.31(6516735-8141017)x1	致病性CNVs, 。包含一个已知致病基因STS基因，该基因与X连锁隐性鱼鳞病相关。	终止妊娠
5	36	arr[hg19]Xp22.31(6516735-8131442)x0	致病性CNVs, 涉及X-连锁隐性遗传鱼鳞病，包含基因STS，主要临床表现为皮肤鱼鳞样改变。	终止妊娠
6	39	arr[hg19]1q21.1q21.2(146501348-148349952)x3	致病性CNVs, 该片段重复涉及1q21.1微重复，其临床表现为自闭症、心脏畸形相关。	终止妊娠
7	38	arr[hg19]Xp11.4p11.23(40871567-46405983)x3	致病性CNVs, 临床表现包括发育迟缓，严重的语言障碍，宽睑裂和鼻孔前倾等。	终止妊娠
8	38	arr[hg19]17q12(34618594-36343793)x3	致病性CNVs, 包含17q12 Recurrent Duplication区域，临床表现智力障碍、语言及运动发育迟缓等。	终止妊娠
9	39	arr[hg19]1q21.1q21.2(146501348-147828939)x1	致病性CNVs, 关键基因为HYDIN2, 临床上有先天性心脏病、孤独症、白内障等表现。	终止妊娠
10	39	arr[hg19]7q11.23(72350815-74138121)x1	致病性CNVs, 包含已知综合征Williams-Beuren综合征区域(WBS)，包含关键基因ELN等，临床表现为心血管畸形，结缔组织异常等。	终止妊娠
11	40	arr[hg19]12p12.3p11.22(14886615-28264747)x1	致病性CNVs，临床上有短指、肌张力降低等，以及发育迟缓, 语言发育障碍，面容异常相关。	终止妊娠
12	38	arr[hg19]15q11.2q13.1(22789021-29050198)x1	致病性CNVs, 包含已知的印记基因SNRPN和UBE3A基因，分别与Prader-Willi综合征(PWS)或者Angelman综合征(AS)相关。	终止妊娠
13	40	arr[hg19]16p13.11(15052746-16303388)x1	致病性CNVs，涉及已知神经认知障碍易感位点，关键基因为NDE1和MYH11，表现为小头畸形，学习障碍，癫痫，行为异常，发育迟缓等。	终止妊娠
14	40	arr[hg19]17p12(14029982-15576592)x1	致病性CNVs，涉及遗传性压迫易感性神经病(herediary neuropathy with liability to pressure palsies，HNPP)，包含关键基因PMP22。临床上有神经受累导致的区域性肌肉无力或麻木等临床特征。	终止妊娠
15	39	arr[hg19]17p12(14101029-15449627)×1	致病性CNVs，涉及已知综合征HNPP，临床表现为压迫后反复出现受累神经支配区域的麻木和肌无力；关键基因为PMP22基因，可导致多种周围性神经病。	终止妊娠
16	40	arr[hg19]22q11.21(18895227-21427379)x1	致病性CNVs, 此区域包含已知综合征(DiGeorge syndrome)，包含关键基因TBX1，此综合征临床表现为心脏异常。	终止妊娠
17	42	arr[hg19]7p22.3p11.2(46239-55716877)x2~3	致病性CNVs，临床表现包括全面发育迟缓、大运动及精细运动发育迟缓、语言发育迟缓、腭裂、房间隔缺损、室中隔缺损、鼻后孔闭锁、大头畸形、癫痫及异常面形等。	终止妊娠
18	41	arr[hg19]16p11.2p12.1(26762221-33918288)x3	致病性CNVs，涉及16p11.2微重复综合征区域，其临床表现为小头畸形、手指脚趾细长、面部畸形、智力低下、婴儿期癫痫等。	终止妊娠
19	37	arr[hg19]Xp22.31(6516735-8131442)x0	致病性CNVs, 涉及Xp22.31缺失区域，包含X-连锁隐性遗传鱼鳞病的关键基因STS。临床表现为皮肤鱼鳞样改变，身材矮小等症状。	终止妊娠
20	37	arr[hg19]Yq11.21q11.221(14687571-15711772)x0	致病性CNVs，涉及Yq11缺失综合征AZFa区域，包含USP9Y基因，与Y染色体连锁精子生成障碍有关，临床表现主要有少(弱)精子症。	继续妊娠
注：CNVs为拷贝数变异；OMIM为在线人类孟德尔遗传数据库；DGV为正常人基因组变异数据。

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