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中华妇幼临床医学杂志(电子版) ›› 2019, Vol. 15 ›› Issue (05) : 541 -546. doi: 10.3877/cma.j.issn.1673-5250.2019.05.010

所属专题: 文献

论著

单核苷酸多态性微阵列芯片技术在胎儿鼻骨缺失检测中的应用
欧阳鲁平1,(), 周桂2, 易赏1, 张月1, 韦慧3, 桂宝恒1, 王锦1   
  1. 1. 广西壮族自治区妇幼保健院遗传代谢中心实验室,南宁 530001
    2. 柳州市工人医院检验科 545005
    3. 广西壮族自治区妇幼保健院超声科,南宁 530001
  • 收稿日期:2019-05-14 修回日期:2019-09-01 出版日期:2019-10-01
  • 通信作者: 欧阳鲁平

Application of single nucleotide polymorphism array technology in genetic examination of nasal bone absence fetuses

Luping Ouyang1,(), Gui Zhou2, Shang Yi1, Yue Zhang1, Hui Wei3, Baoheng Gui1, Jin Wang1   

  1. 1. Laboratory of Genetic and Metabolism Center, Maternal & Child Health Hospital of Guangxi Zhuang Autonomous Region, Nanning 530001, Guangxi Zhuang Autonomous Region, China
    2. Clinical Laboratory of Liuzhou Workers Hospital, Liuzhou 545005, Guangxi Zhuang Autonomous Region, China
    3. Department of Ultround, Maternal & Child Health Hospital of Guangxi Zhuang Autonomous Region, Nanning 530001, Guangxi Zhuang Autonomous Region, China
  • Received:2019-05-14 Revised:2019-09-01 Published:2019-10-01
  • Corresponding author: Luping Ouyang
  • About author:
    Corresponding author: Ouyang Luping, Email:
  • Supported by:
    National Natural Science Foundation of China(2018JJA140303); Natural Science Foundation of Guangxi Zhuang Autonomous Region(2016JJB140208); Self-Raised Project of Health Commission of Guangxi Zhuang Autonomous Region(Z20190054)
引用本文:

欧阳鲁平, 周桂, 易赏, 张月, 韦慧, 桂宝恒, 王锦. 单核苷酸多态性微阵列芯片技术在胎儿鼻骨缺失检测中的应用[J]. 中华妇幼临床医学杂志(电子版), 2019, 15(05): 541-546.

Luping Ouyang, Gui Zhou, Shang Yi, Yue Zhang, Hui Wei, Baoheng Gui, Jin Wang. Application of single nucleotide polymorphism array technology in genetic examination of nasal bone absence fetuses[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2019, 15(05): 541-546.

目的

探讨染色体核型分析与单核苷酸多态性微阵列芯片(SNP-array)检查在胎儿鼻骨缺失遗传学检查中的应用价值。

方法

选取2016年1月至2018年10月,于广西壮族自治区妇幼保健院进行产前检查,并且超声检查结果提示胎儿鼻骨缺失的241例胎儿及其母亲为研究对象。回顾性分析241例胎儿鼻骨缺失的孕妇分别于早、中和晚孕期分别行介入性产前诊断绒毛、羊水、脐带血标本的临床资料,同时行染色体核型分析与SNP-array检查。SNP-array检查采Illumina Human Cyto SNP12微阵列芯片进行全基因组拷贝数变异(CNVs)检测,结合查询国际病理性CNVs数据库(ClinGen、ClinVar、DECIPHER、OMIM),正常人基因组变异数据库(DGV)以及PubMed文献数据库等对检出的CNVs的致病性进行分析。本研究经广西壮族自治区妇幼保健院伦理委员会批准[医研伦快审(2019)第(3-20)号],并与受试者监护人签署临床研究知情同意书。

结果

241例胎儿鼻骨缺失孕妇的绒毛、羊水及脐血样本中,检出染色体核型异常为30例,异常检出率12.4%(30/241);SNP-array检出异常为43例,异常检出率为17.8%(43/241)。染色体核型检出的30例产前诊断标本异常核型如下:24例(80.0%)为21-三体综合征,3例(10.0%)为18-三体综合征,2例(6.7%)为13-三体综合征,以及1例(3.3%)为平衡易位。染色体正常但是SNP-array检出异常者中,3例为致病性CNVs,1例疑似为致病性CNVs,以及10例为临床意义不明CNVs。

结论

对超声检测提示胎儿鼻骨缺失的胎儿样本,行SNP-array检测有助于发现染色体核型分析无法检出的染色体亚显微结构异常,并且SNP-array有利于提高对胎儿鼻骨缺失的遗传病因的诊断。

Objective

To evaluate the application value of chromosome karyotype analysis and single nucleotide polymorphism array (SNP-array) technology in genetic examination of nasal bone absence fetuses.

Methods

From January 2016 to October 2018, 241 fetuses and their mothers were selected for prenatal examination in Maternal & Child Health Hospital of Guangxi Zhuang Autonomous Region, and the 241 fetuses were diagnosed as nasal bone absence by the results of ultrasound examination. The clinical data of 241 cases were retrospectively analyzed. Interventional prenatal diagnosis of villi, amniotic fluid and umbilical cord blood samples were performed in first trimester, second trimester and third trimester, respectively. Chromosome karyotype analysis and SNP-array examination were performed at the same time. SNP-array examination adopts Illumina Human Cyto SNP12 microarray chip to carry out whole genome copy number variation (CNVs) detection. The pathogenicity of CNVs detected is analyzed by querying international pathological CNVs database (ClinGen, ClinVar, DECIPHER, OMIM), normal human genome variation database (DGV) and PubMed literature database. This study was approved by the ethics committee of Maternal & Child Health Hospital of Guangxi Zhuang Autonomous Region [2019 No. (3-20)]. Informed consent was obtained from each participate′s guardian.

Results

In 241 cases of fetal samples with flank, amniotic fluid and umbilical cord blood, the karyotype was detected in 30 cases, the detection rate was 12.4% (30/241), and SNP-array detected 43 cases with abnormal detection rate of 17.8% (43/241). The abnormal karyotypes of 30 cases of prenatal diagnosis were as follows: 24 cases (80.0%) of 21-trisomy syndrome, 3 cases(10.0%) of 18-trisomy syndrome, 2 cases (6.7%) of 13-trisomy syndrome and 1 case (3.3%) of balanced translocation. Among those with normal chromosomes but abnormal chips, 3 cases were pathogenic CNVs, 1 case was suspected to be pathogenic CNVs, and 10 cases were clinically unknown CNVs.

Conclusions

SNP-array detection with fetal nasal bone absence by ultrasound can help to detect chromosome submicroscopic structural abnormalities that cannot be detected by karyotype analysis, and SNP-array is beneficial for improving the diagnosis of genetic causes of fetal nasal bone absence.

表1 鼻骨缺失胎儿标本的SNP-array检测与染色体核型分析情况比较(例)
表2 4例鼻骨缺失胎儿中的致病CNVs或疑似致病性CNVs
病例 标本类型 产前诊断指征 检出拷贝数变异 数据库查询结果 妊娠结局
1 羊水 胎儿鼻骨缺失、高龄妊娠 arr (22)x2~3,嵌合 致病性CNVs。羊水细胞核型检测22号染色体三体嵌合比例为4%~35%的患者亦可表现出发育迟缓,特殊面容,皮质萎缩,肌张力降低,先天性心脏系统异常等1种或多种临床症状 终止妊娠
2 脐血 胎儿鼻骨缺失 arr 8p23.1(8101641-12690464)x3,4.5 Mb重复 致病性CNVs。该重复区域涉及8p23.1重复综合征,包含关键基因GATA4等,8p23.1重复综合征患者常见的临床表现宽泛,包括轻至中度发育迟缓,学习困难,语言发育迟缓,行为困难,面容异常,视力异常,心血管异常,大头畸形,注意力障碍等,在患者数据库ClinVar、Decipher中有大量致病性报道,患者临床表现包括面容异常,发育迟缓,智力障碍,心血管异常等 终止妊娠
3 羊水 鼻骨缺失、高龄妊娠、左心室强光斑 arr17p12(14520034-15160859)x1, 0.64 Mb的片段缺失 致病性CNVs。此缺失位于已知综合征HNPP内,并且包含关键基因PMP22部分区域,该基因是剂量敏感性基因,HNPP的临床表现为反复发作的、易受嵌压部位的周围神经麻痹,包括臂丛或腓总神经、尺骨神经、正中神经等,表现为受累神经支配区的深浅感觉减退、所支配的肌肉张力减退,甚至会出现肌肉萎缩、关节屈曲挛缩。约10%患者具有高足弓及槌状趾 终止妊娠
4 脐血 胎儿左侧鼻骨缺失并右侧鼻骨发育不良、高龄妊娠 arr2q13(111392259-113212450)x1,1.8 Mb的片段缺失 疑似致病性CNVs。2号染色体该片段缺失在ClinVar及Decipher数据库中可见数例致病性的文献报道。患者临床表现包括颅骨异常、自闭症、轻度智力障碍、面神经麻痹、语言发育迟缓等。文献报道,2q13缺失的患者临床异质性较大并且外显不全,共同临床表现包括发育迟缓及轻微的颅面畸形等[10]。该片段包含2个已知OMIM致病基因,该片段缺失在普通人数据库DGV中有2例文献报道,为疑似致病性CNVs 终止妊娠
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