Gunnarsson Rebeqa, Staaf Johan, Jansson Mattias, Ottesen Anne Marie, Göransson Hanna, Liljedahl Ulrika, Ralfkiaer Ulrik, Mansouri Mahmoud, Buhl Anne Mette, Smedby Karin Ekström, Hjalgrim Henrik, Syvänen Ann-Christine, Borg Ake, Isaksson Anders, Jurlander Jesper, Juliusson Gunnar, Rosenquist Richard
Lund Strategic Research Center for Stem Cell Biology and Cell Therapy, Hematology and Transplantation, Lund University, Lund, Sweden.
Genes Chromosomes Cancer. 2008 Aug;47(8):697-711. doi: 10.1002/gcc.20575.
Screening for gene copy-number alterations (CNAs) has improved by applying genome-wide microarrays, where SNP arrays also allow analysis of loss of heterozygozity (LOH). We here analyzed 10 chronic lymphocytic leukemia (CLL) samples using four different high-resolution platforms: BAC arrays (32K), oligonucleotide arrays (185K, Agilent), and two SNP arrays (250K, Affymetrix and 317K, Illumina). Cross-platform comparison revealed 29 concordantly detected CNAs, including known recurrent alterations, which confirmed that all platforms are powerful tools when screening for large aberrations. However, detection of 32 additional regions present in 2-3 platforms illustrated a discrepancy in detection of small CNAs, which often involved reported copy-number variations. LOH analysis using dChip revealed concordance of mainly large regions, but showed numerous, small nonoverlapping regions and LOH escaping detection. Evaluation of baseline variation and copy-number ratio response showed the best performance for the Agilent platform and confirmed the robustness of BAC arrays. Accordingly, these platforms demonstrated a higher degree of platform-specific CNAs. The SNP arrays displayed higher technical variation, although this was compensated by high density of elements. Affymetrix detected a higher degree of CNAs compared to Illumina, while the latter showed a lower noise level and higher detection rate in the LOH analysis. Large-scale studies of genomic aberrations are now feasible, but new tools for LOH analysis are requested.
通过应用全基因组微阵列,基因拷贝数改变(CNA)的筛查得到了改进,其中单核苷酸多态性(SNP)阵列还允许分析杂合性缺失(LOH)。我们在此使用四种不同的高分辨率平台分析了10例慢性淋巴细胞白血病(CLL)样本:细菌人工染色体(BAC)阵列(32K)、寡核苷酸阵列(185K,安捷伦)以及两种SNP阵列(250K,Affymetrix和317K,Illumina)。跨平台比较发现了29个一致检测到的CNA,包括已知的复发性改变,这证实了在筛查大片段畸变时所有平台都是强大的工具。然而,在2 - 3个平台中检测到的另外32个区域表明,在检测小CNA时存在差异,这些小CNA往往涉及已报道的拷贝数变异。使用dChip进行的LOH分析显示主要是大片段区域的一致性,但也显示出许多小的不重叠区域以及未被检测到的LOH。对基线变异和拷贝数比反应的评估显示安捷伦平台表现最佳,并证实了BAC阵列的稳健性。因此,这些平台显示出更高程度的平台特异性CNA。SNP阵列显示出更高的技术变异,尽管这被元件的高密度所补偿。与Illumina相比,Affymetrix检测到的CNA程度更高,而后者在LOH分析中显示出较低的噪声水平和较高的检测率。现在对基因组畸变进行大规模研究是可行的,但需要新的LOH分析工具。