Boone Elke, Heezen Kim C, Groenen Patricia J T A, Langerak Anton W
Laboratory for Molecular Diagnostics, Department of Laboratory Medicine, AZ Delta Hospital, Roeselare, Belgium.
Laboratory Medical Immunology, Department of Immunology, Erasmus MC, Rotterdam, Netherlands.
Methods Mol Biol. 2019;1956:77-103. doi: 10.1007/978-1-4939-9151-8_4.
Assessment of the presence of clonal lymphoproliferations via polymerase chain reaction (PCR)-based analysis of rearranged immunoglobulin (IG) or T-cell receptor (TR) genes is a valuable method in the diagnosis of suspect lymphoproliferative disorders. Additionally, this methodology can be used for evaluating dissemination of lymphoma cells and for studying the clonal relationship between multiple (different locations) or consecutive (over time) lymphomas. Here we describe an integrated approach to assess clonality via analysis of Ig heavy chain (IGH), Ig kappa (IGK), TCR beta (TRB), and TCR gamma (TRG) gene rearrangements, based on the standardized multiplex PCRs as originally developed by the European BIOMED-2 consortium. The described protocol covers the pre-analytical phase of DNA isolation (from formalin-fixed paraffin-embedded and fresh tissues, body fluids, peripheral blood, and bone marrow), the analytical phase of PCR GeneScan and heteroduplex analysis, and the post-analytical interpretation of the obtained profiles, following established guidelines.
通过基于聚合酶链反应（PCR）的重排免疫球蛋白（IG）或T细胞受体（TR）基因分析来评估克隆性淋巴细胞增殖的存在，是诊断可疑淋巴细胞增殖性疾病的一种有价值的方法。此外，该方法可用于评估淋巴瘤细胞的播散情况，并研究多个（不同部位）或连续（随时间）淋巴瘤之间的克隆关系。在此，我们描述一种基于欧洲BIOMED-2联盟最初开发的标准化多重PCR，通过分析Ig重链（IGH）、Igκ（IGK）、TCRβ（TRB）和TCRγ（TRG）基因重排来评估克隆性的综合方法。所描述的方案涵盖了DNA分离的分析前阶段（从福尔马林固定石蜡包埋组织和新鲜组织、体液、外周血和骨髓中提取）、PCR基因扫描和异源双链分析的分析阶段，以及按照既定指南对获得的图谱进行分析后解释。
