Brinckmann R, Kaufmann O, Reinartz B, Dietel M
Institute of Pathology, Charité University Hospital, Berlin, Germany.
J Pathol. 2000 Jan;190(1):55-60. doi: 10.1002/(SICI)1096-9896(200001)190:1<55::AID-PATH501>3.0.CO;2-1.
A study was performed to investigate the utility of polymerase chain reaction (PCR)-based analysis of immunoglobulin heavy chain (IgH) gene rearrangements for the diagnosis of low-grade malignant B-cell lymphomas on formalin-fixed, EDTA-decalcified, and paraffin-embedded bone marrow trephine biopsies. On amplifying two DNA samples per biopsy, no reproducible monoclonal PCR result was found in 32 patients with reactive lymphoid hyperplasias. In contrast, 5/14 patients with known low-grade B-cell lymphomas, but histomorphologically and immunohistochemically lymphoma-free bone marrow, showed a reproducible monoclonal IgH gene rearrangement. In three of these cases, sequence analysis revealed completely different amplification products on comparing bone marrow and lymph node infiltrations, while in the other two cases the products were identical. In one of the latter biopsies, an unequivocal lymphoma infiltrate was found after step sectioning of the biopsy, while the other case remained lymphoma-free according to conventional criteria. A third group of three patients with known lymphomas and bone marrow findings that were suggestive but not diagnostic of bone marrow involvement showed monoclonal PCR results in all three cases, with identical sequences in bone marrow and extramedullary lymphoma infiltrates. These data suggest that a reproducible monoclonal IgH gene rearrangement is highly specific for the presence of malignant B-cells in bone marrow. In staging procedures for low-grade B-cell lymphomas, PCR yields no additional information in cases that are morphologically and immunohistochemically lymphoma-free after evaluation of representative sections. PCR may be useful in equivocal cases, provided that IgH gene rearrangements of extramedullary lymphoma and bone marrow are sequenced and compared.
进行了一项研究，以调查基于聚合酶链反应（PCR）分析免疫球蛋白重链（IgH）基因重排，在福尔马林固定、乙二胺四乙酸（EDTA）脱钙和石蜡包埋的骨髓活检组织中诊断低度恶性B细胞淋巴瘤的效用。在每份活检组织扩增两个DNA样本时，32例反应性淋巴组织增生患者未发现可重复的单克隆PCR结果。相比之下，14例已知低度B细胞淋巴瘤患者中，有5例骨髓组织形态学和免疫组织化学检查未见淋巴瘤，但显示出可重复的单克隆IgH基因重排。其中3例，序列分析显示骨髓浸润和淋巴结浸润的扩增产物完全不同，而另外2例产物相同。在后2例活检组织中，其中1例在活检组织连续切片后发现明确的淋巴瘤浸润，而另1例根据传统标准仍未见淋巴瘤。第三组3例已知淋巴瘤患者，骨髓检查结果提示但不能确诊骨髓受累，所有3例均显示单克隆PCR结果，骨髓浸润和髓外淋巴瘤浸润序列相同。这些数据表明，可重复的单克隆IgH基因重排对骨髓中恶性B细胞的存在具有高度特异性。在低度B细胞淋巴瘤分期过程中，对于评估代表性切片后形态学和免疫组织化学检查未见淋巴瘤的病例，PCR未提供额外信息。在可疑病例中，PCR可能有用，前提是对髓外淋巴瘤和骨髓的IgH基因重排进行测序和比较。
