Konialis Christopher, Savola Suvi, Karapanou Sophia, Markaki Aggeliki, Karabela Maria, Polychronopoulou Sophia, Ampatzidou Maria, Voulgarelis Michael, Viniou Nora-Athina, Variami Eleni, Koumarianou Argyro, Zoi Katerina, Hagnefelt Birgitta, Schouten Jan P, Pangalos Constantinos
Hematology. 2014 Jun;19(4):217-24. doi: 10.1179/1607845413Y.0000000112. Epub 2013 Nov 25.
The presence of numerical and/or structural chromosomal abnormalities is a frequent finding in clonal hematopoietic malignant disease, typically diagnosed through routine karyotyping and/or fluorescent in situ hybridization (FISH) analysis. Recently, the application of array comparative genomic hybridization (aCGH) has uncovered many new cryptic genomic copy number imbalances, most of which are now recognized as clinically useful markers of haematological malignancies. In view of the limitations of both FISH and aCGH techniques, in terms of their routine application as a first line screening test, we designed a new multiple ligation-dependent probe amplification (MLPA) probemix for use in addition to classic karyotype analysis.
A novel MLPA probemix was developed to interrogate copy number changes involving chromosomal regions: 2p23-24 (MYCN, ALK), 5q32-34 (MIR145A, EBF1, MIR146A), 6q21-27, 7p12.2 (IKZF1), 7q21-36, 8q24.21 (MYC), 9p24 (JAK2 V617F point mutation), 9p21.3 (CDKN2A/2B), 9p13.2 (PAX5), 10q23 (PTEN), 11q22.3 (ATM), 12p13.2 (ETV6), 13q14 (RB1, MIR15A, DLEU2, DLEU1), 17p13.1 (TP53), and 21q22.1 (RUNX1/AML1) and was applied to DNA extracted from 313 consecutive bone marrow patient samples, referred for routine karyotype analysis.
More than half of the samples originated from newly investigated patients. We discovered clinically relevant genomic aberrations, involving a total of 24 patients (8%) all with a normal karyotype, which would have remained undiagnosed.
Our data clearly indicate that routine application of this MLPA screening panel, as an adjunct to karyotype analysis, provides a sensitive, robust, rapid and low-cost approach for uncovering clinically important genomic abnormalities, which would have otherwise remained undetected.
在克隆性造血恶性疾病中，经常会发现染色体数目和/或结构异常，通常通过常规核型分析和/或荧光原位杂交（FISH）分析来诊断。最近，阵列比较基因组杂交（aCGH）的应用发现了许多新的隐匿性基因组拷贝数失衡，其中大多数现在被认为是血液系统恶性肿瘤的临床有用标志物。鉴于FISH和aCGH技术在作为一线筛查试验的常规应用方面存在局限性，我们设计了一种新的多重连接依赖探针扩增（MLPA）探针混合物，并将其与经典核型分析一起使用。
开发了一种新型MLPA探针混合物，用于检测涉及以下染色体区域的拷贝数变化：2p23 - 24（MYCN、ALK）、5q32 - 34（MIR145A、EBF1、MIR146A）、6q21 - 27、7p12.2（IKZF1）、7q2l - 36、8q24.21（MYC）、9p24（JAK2 V617F点突变）、9p21.3（CDKN2A/2B）、9p13.2（PAX5）、10q23（PTEN）、11q22.3（ATM）、12p13.2（ETV6）、13q14（RB1、MIR15A、DLEU2、DLEU1）、17p13.1（TP53）和21q22.1（RUNX1/AML1），并将其应用于从313例连续骨髓患者样本中提取的DNA，这些样本被送去进行常规核型分析。
超过一半的样本来自新接受检查的患者。我们发现了临床上相关的基因组畸变，共有24例患者（8%）存在这种情况，他们的核型均正常，若不进行此项检测这些畸变将无法被诊断出来。
我们的数据清楚地表明，作为核型分析的辅助手段，常规应用这种MLPA筛查面板能够提供一种灵敏、可靠、快速且低成本的方法，用于发现临床上重要的基因组异常，否则这些异常将无法被检测到。
