Dawidowska Małgorzata, Derwich Katarzyna, Szczepański Tomasz, Jółkowska Justyna, Witt Michał, Wachowiak Jacek
Zakład Genetyki Molekularnej i Klinicznej, Instytut Genetyki Człowieka PAN, ul. Strzeszyńska 32, 60-479 Poznań, Poland.
Med Wieku Rozwoj. 2006 Jan-Mar;10(1 Pt 2):323-34.
Initiation and popularization of routine molecular diagnostics of minimal residual disease (MRD) are currently one of the most urgent challenges in Polish hemato-oncology. The paper is aimed to present preliminary results of identification of immunoglobulin (Ig) and T-cell receptor (TCR) gene rearrangements and quantitative assessment of MRD levels in Polish children with acute lymphoblastic leukemia (ALL). The results are presented in the context of clinical significance of MRD study, current methodology of MRD assessment and standardization process in Western Europe.
DNA isolated from bone marrow / bone marrow mononuclear cells obtained at diagnosis from 26 children (25 B-precursor ALL, 1 T-ALL) aged 1.3-16.5 years.
PCR-heteroduplex analysis, based on standard BIOMED-1 and BIOMED-2 primer combinations and protocols for detection of rearrangements and clonality assessment; sequencing of clonal PCR products and comparison with germline sequences of Ig/TCR genes for identification of the rearranged genes andjunctional regions; real-time quantitative PCR (RQ-PCR) with the use of TaqMan probes for assessment of follow-up MRD levels (in 11 patients).
Clonal TCRG, incomplete TCRD, Vdelta2-Jalpha, TCRB, IGK-Kde and IGH gene rearrangements were detected in 61, 61, 35, 13, 39 and 83% of patients, respectively, which was generally concordant with published data for patients of other European nations.
There is an urgent need to broaden the scope of minimal residual disease study in Poland and to develop Polish standards of MRD diagnostics, based on current European experience and standards.
启动并推广微小残留病（MRD）的常规分子诊断，是目前波兰血液肿瘤学中最紧迫的挑战之一。本文旨在介绍波兰急性淋巴细胞白血病（ALL）患儿免疫球蛋白（Ig）和T细胞受体（TCR）基因重排的鉴定以及MRD水平定量评估的初步结果。研究结果是在MRD研究的临床意义、当前MRD评估方法以及西欧标准化进程的背景下呈现的。
从26名年龄在1.3 - 16.5岁儿童（25例B前体ALL，1例T - ALL）诊断时获取的骨髓/骨髓单个核细胞中分离的DNA。
基于标准的BIOMED - 1和BIOMED - 2引物组合及方案进行PCR - 异源双链分析，用于检测重排和克隆性评估；对克隆性PCR产物进行测序，并与Ig/TCR基因的种系序列比较，以鉴定重排基因和连接区；使用TaqMan探针进行实时定量PCR（RQ - PCR），以评估随访时的MRD水平（11例患者）。
分别在61%、61%、35%、13%、39%和83%的患者中检测到克隆性TCRG、不完全TCRD、Vdelta2 - Jalpha、TCRB、IGK - Kde和IGH基因重排，这与其他欧洲国家患者的已发表数据总体一致。
基于当前欧洲的经验和标准，波兰迫切需要扩大微小残留病研究的范围，并制定波兰的MRD诊断标准。
