Scott Daryl A, Klaassens Merel, Holder Ashley M, Lally Kevin P, Fernandes Caraciolo J, Galjaard Robert-Jan, Tibboel Dick, de Klein Annelies, Lee Brendan
Department of Molecular and Human Genetics, Baylor College of Medicine, One Baylor Plaza, Houston, TX 77030, USA.
Hum Mol Genet. 2007 Feb 15;16(4):424-30. doi: 10.1093/hmg/ddl475. Epub 2007 Jan 8.
Non-isolated congenital diaphragmatic hernia (CDH+) is a severe birth defect that is often caused by de novo chromosomal anomalies. In this report, we use genome-wide oligonucleotide-based array comparative genome hybridization (aCGH) followed by rapid real-time quantitative PCR analysis to identify, confirm and map chromosomal anomalies in a cohort of 26 CDH+ patients. One hundred and five putative copy number changes were identified by aCGH in our cohort of CDH+ patients. Sixty-one of these changes (58%) had been previously described in normal controls. Twenty of the remaining 44 changes (45%) were confirmed by quantitative real-time PCR or standard cytogenetic techniques. These changes included de novo chromosomal abnormalities in five of the 26 patients (19%), two of whom had previously normal G-banded chromosome analyses. Data from these patients provide evidence for the existence of CDH-related genes on chromosomes 2q37, 6p22-25 and 14q, and refine the CDH minimal deleted region on 15q26 to an interval that contains COUP-TFII and only eight other known genes. Although COUP-TFII is likely to play a role in the development of CDH in patients with 15q26 deletions, we did not find COUP-TFII mutations in 73 CDH samples. We conclude that the combination of oligonucleotide-based aCGH and quantitative real-time PCR is an effective method of identifying, confirming and mapping clinically relevant copy number changes in patients with CDH+. This method is more sensitive than G-banded chromosome analysis and may find wide application in screening patients with congenital anomalies.
非孤立性先天性膈疝（CDH+）是一种严重的出生缺陷，通常由新发染色体异常引起。在本报告中，我们使用基于全基因组寡核苷酸的阵列比较基因组杂交（aCGH），随后进行快速实时定量PCR分析，以识别、确认和定位26例CDH+患者队列中的染色体异常。通过aCGH在我们的CDH+患者队列中鉴定出105个假定的拷贝数变化。这些变化中的61个（58%）先前已在正常对照中描述过。其余44个变化中的20个（45%）通过定量实时PCR或标准细胞遗传学技术得到确认。这些变化包括26例患者中的5例（19%）存在新发染色体异常，其中2例先前G带染色体分析正常。这些患者的数据为2q37、6p22 - 25和14号染色体上存在CDH相关基因提供了证据，并将15q26上的CDH最小缺失区域细化为一个包含COUP - TFII和仅其他8个已知基因的区间。尽管COUP - TFII可能在15q26缺失患者的CDH发生中起作用，但我们在73个CDH样本中未发现COUP - TFII突变。我们得出结论，基于寡核苷酸的aCGH和定量实时PCR的组合是识别、确认和定位CDH+患者临床相关拷贝数变化的有效方法。该方法比G带染色体分析更敏感，可能在先天性异常患者的筛查中得到广泛应用。
