The diagnosis and risk stratification of multiple myeloma (MM) is based on clinical and cytogenetic tests. Magnetic CD138 enrichment followed by interphase FISH (fluorescence in situ hybridisation) is the gold standard to identify prognostic translocations and copy number alterations (CNA). Although clinical implications of gene expression profiling (GEP) or panel based sequencing results are evident, those tests have not yet reached routine clinical application. We set up a single workflow to analyse MM of 211 patients at first diagnosis by whole genome sequencing (WGS) and RNA-Seq and validate the results by FISH analysis. We observed a 96% concordance of FISH and WGS results when assessing translocations involving the IGH locus and an overall concordance of FISH and WGS of 92% when assessing CNA. WGS analysis resulted in the identification of 17 additional MYC-translocations that were missed by FISH analysis. RNA-Seq followed by supervised clustering grouped patients in their expected genetically defined subgroup and prompted the assessment of WGS data in cases that were not congruent with FISH. This allowed the identification of additional IGH-translocations and hyperdiploid cases. We show the reliability of WGS an RNA-Seq in a clinical setting, which is a prerequisite for a novel routine diagnostic test.

多发性骨髓瘤（MM）的诊断和风险分层基于临床和细胞遗传学检测。磁性CD138富集，然后进行相间FISH（荧光原位杂交）是鉴定预后易位和拷贝数改变（CNA）的金标准。尽管基因表达谱（GEP）或基于面板的测序结果的临床意义很明显，但这些测试尚未达到常规临床应用。我们建立了一个单一的工作流程，通过全基因组测序（WGS）和RNA-Seq对初诊时211例患者的MM进行分析，并通过FISH分析验证结果。在评估涉及IGH基因座的易位时，我们观察到FISH和WGS结果的一致性为96％，而在评估CNA时，FISH和WGS的总体一致性为92％。FISH分析遗漏的MYC易位。RNA-Seq随后是有监督的聚类，将患者分组在其预期的遗传定义亚组中，并在与FISH不符的病例中提示对WGS数据进行评估。这样可以鉴定其他IGH易位和超二倍体病例。我们在临床环境中显示了WGS RNA-Seq的可靠性，这是进行新型常规诊断测试的前提。