多重连接探针扩增技术在先天性心脏病22q11微缺失/微重复综合征诊断中的应用

杨月华; 胡娅莉; 朱湘玉; 莫绪明; 王东进; 姚金翠; 盛敏; 朱海燕; 李洁; 茹彤; 王志群

PDF(1069 KB)

中国当代儿科杂志 ›› 2009, Vol. 11 ›› Issue (12) : 892-896.

论著·临床研究

多重连接探针扩增技术在先天性心脏病22q11微缺失/微重复综合征诊断中的应用

杨月华，胡娅莉，朱湘玉，莫绪明，王东进，姚金翠，盛敏，朱海燕，李洁，茹彤，王志群

作者信息 +

Diagnosis of 22q11 deletion and duplication in congenital heart disease by multiplex ligation-dependent probe amplification

YANG Yue-Hua, HU Ya-Li, ZHU Xiang-Yu, MO Xu-Ming, WANG Dong-Jin, YAO Jin-Cui, SHENG Min, ZHU Hai-Yan, LI Jie, RU Tong, WANG Zhi-Qun

Author information +

文章历史 +

摘要

目的:染色体22q11区域基因拷贝数异常是先天性心脏病（CHD）的遗传病因之一，由其引起的CHD预后不良。该研究主要探讨多重连接探针扩增技术用于CHD 22q11微缺失或微重复遗传病因诊断的实用性，并了解22q11微缺失或微重复在CHD中的发生情况。方法:选择25个位于染色体22q11低重复拷贝序列A-H区域内、7个位于其周围（CES、22q13）和16个位于4、8、10、17号染色体上的基因位点共计48个探针组成多重连接探针，对181例外科手术前的CHD儿童和14例严重CHD或包括CHD的多发性畸形胎儿进行了22q11微缺失或微重复的检测，并进行了染色体核型分析。结果:195例患儿中，共检出22q11微缺失者7例（LCR A-D区6例，LCR A-C区1例），22q11微重复1例（LCR B-D区），涉及的CHD类型包括室间隔缺损、房室间隔缺损、肺动脉狭窄和法洛四联征。同时染色体核型分析还发现了6例异常：1例21q部分缺失［46，XY，21q-］，1例嵌合性8-三体［47，XY，+8/46，XY（1∶2）］，4例21-三体。其中1例21-三体与22q11微重复同时存在。结论:染色体22q11区域高密度多重连接探针检测技术能快速、灵敏、精确定位诊断染色体22q11区域基因拷贝数异常，适合于CHD的遗传学诊断；此外，22q11微缺失或微重复引起的CHD类型多种多样，建议所有CHD患者应常规进行遗传学检测。［中国当代儿科杂志，2009，11（11）：892-896］

Abstract

OBJECTIVE: To investigate the clinical utility of multiplex ligation-dependent probe amplification (MLPA) for detecting 22q11 deletion and duplication in congenital heart disease (CHD) cases and to study the incidence of 22q11 deletion and duplicaton in different kinds of CHD. METHODS: Forty-eight probes of which 25 located in 22q11 low copy number region (LCR 22s A-H), 7 in 22q11 surrounding region (CES, 22q13) and 16 in chromosomes 4, 8, 10 and 17 were selected to detect 22q11 deletion and duplication in 181 preoperative children with CHD and 14 fetuses with serious CHD or CHD with multiple malformations. In these cases, karyotype analysis was also performed. RESULTS: MLPA demonstrated that 7 cases had 22q11 deletion ［6 cases from CLTCL1 to LZTR1(LCR A-D) and 1 case from CLTCL1 to PCQAP (LCR A-C)］ and that 1 case had 22q11 duplication，spanning from ZNF74 to LZTR1(LCR B-D). The phenotypes of heart defect included ventricular septal defect, atrioventricular septal defect, pulmonary stenosis and tetralogy of Fallot. Karyotype analysis showed that 1 case had 21q deletion ［46, XY, 21q］, 1 case had mosaic trisomy 8 ［ 47,XY,+8/46, XY(1∶2)］ and 4 cases had trisomy 21. One of the 4 cases with trisomy 21 had concurrent 22q11 duplication. CONCLUSIONS: MLPA is a rapid, sensitive, site-specific and relatively inexpensive method for diagnosis of 22q11 deletion and duplication in CHD. 22q11 deletion and duplication may cause various kinds of CHD, suggesting that genetic detection should be performed routinely in CHD patients.［Chin J Contemp Pediatr, 2009, 11 (11):892-896］

导出引用

杨月华, 胡娅莉, 朱湘玉, 莫绪明, 王东进, 姚金翠, 盛敏, 朱海燕, 李洁, 茹彤, 王志群. 多重连接探针扩增技术在先天性心脏病22q11微缺失/微重复综合征诊断中的应用[J]. 中国当代儿科杂志. 2009, 11(12): 892-896

YANG Yue-Hua, HU Ya-Li, ZHU Xiang-Yu, MO Xu-Ming, WANG Dong-Jin, YAO Jin-Cui, SHENG Min, ZHU Hai-Yan, LI Jie, RU Tong, WANG Zhi-Qun. Diagnosis of 22q11 deletion and duplication in congenital heart disease by multiplex ligation-dependent probe amplification[J]. Chinese Journal of Contemporary Pediatrics. 2009, 11(12): 892-896

中图分类号： R541.1 R446

参考文献

［1］Botto LD, May K, Fernhoff PM, Correa A, Coleman K, Rasmussen SA, et al. A population-based study of the 22q11.2 deletion: phenotype, incidence, and contribution to major birth defects in the population［J］. Pediatrics, 2003, 112（1 Pt 1）：101-107.
［2］Kyburz A, Bauersfeld U, Schinzel A, Riegel M, Hug M, Tomaske M, et al.The fate of children with microdeletion 22q11.2 syndrome and congenital heart defect：clinical course and cardiac outcome［J］. Pediatr Cardiol, 2008, 29(1)：76-83.
［3］Park IS, Ko JK, Kim YH, Yoo HW, Seo EJ, Choi JY, et al. Cardiovascular anomalies in patients with chromosome 22q11.2 deletion：a Korean multicenter study［J］. Int J Cardiol, 2007, 114 (2)：230-235.
［4］许争峰，易龙，莫绪明，胡娅莉，王东进，朱瑞芳，等. 先天性心脏病患者22q11 微缺失检测及相关分析［J］. 中华医学遗传学杂志，2006，23(3)：250-255.
［5］许争峰，胡娅莉，张建伟，朱瑞芳，茹彤，刘启兰，等. 9 例18 三体综合征胎儿的产前诊断［J］. 中华围产医学杂志，2006，9 (5)：291-293.
［6］郑明明，胡娅莉，许争峰，王志群，史婷琦，叶晓东，等.实时荧光定量聚合酶链反应技术在产前诊断唐氏综合征中的应用［J］. 中华妇产科杂志，2004，39(10)：678-681.
［7］Jalali GR, Vorstman JA, Errami A, Vijzelaar R, Biegel J, Shaikh T, et al. Detailed analysis of 22q11.2 with a high density MLPA probe set［J］. Hum Mutat, 2008, 29(3)：433-440.
［8］Schouten JP, McElgunn CJ, Waaijer R, Zwijnenburg D, Diepvens F, Pals G. Relative quantification of 40 nucleic acid sequences by multiplex ligation-dependent probe amplification［J］. Nucleic Acids Res, 2002, 30(12)：e57.
［9］Fernández L, Lapunzina P, Arjona D, López Pajares I, García-Guereta L, Elorza D, et al. Comparative study of three diagnostic approaches (FISH, STRs and MLPA) in 30 patients with 22q 11.2 deletion syndrome［J］. Clin Genet, 2005, 68(4)：373-378.
［10］Stachon AC, Baskin B, Smith AC, Shugar A, Cytrynbaum C, Fishman L, et al. Molecular diagnosis of 22q11.2 deletion and duplication by multiplex ligation dependent probe amplification［J］. Am J Med Genet A, 2007,143A(24)：2924-2930.
［11］Shaikh TH, O′Connor RJ, Pierpont ME, McGrath J, Hacker AM, Nimmakayalu M, et al. Low copy repeats mediate distal chromosome 22q11.2 deletions： sequence analysis predicts breakpoint mechanisms［J］.Genome Res, 2007, 17(4)：482-491.
［12］Courtens W, Schramme I, Laridon A. Microduplication 22q11.2：a benign polymorphism or a syndrome with a very large clinical variability and reduced penetrance?—Report of two families［J］. Am J Med Genet A, 2008, 146A(6)：758-763.