Diss T C, Peng H, Wotherspoon A C, Isaacson P G, Pan L
Department of Histopathology, University College London Medical School, U.K.
J Pathol. 1993 Mar;169(3):291-5. doi: 10.1002/path.1711690303.
Detection of B-cell monoclonality using the polymerase chain reaction (PCR) promises the quick and cost-effective separation of monoclonal from polyclonal B-cell disease. However, the efficiency of the method has yet to be fully assessed, particularly with regard to disease type and selection of PCR primers. We have evaluated two approaches based on amplification of the immunoglobulin heavy chain gene using framework 2 (Fr2) and framework 3 (Fr3) region primers. Frozen tissue samples from 94 cases of low-grade B-cell lymphoma were investigated, all of which had previously been shown to be monoclonal by Southern blot analysis. Using a Fr2 primer, we were able to show monoclonality in 85 per cent of cases; with Fr3, 80 per cent of cases; and using both techniques in separate reactions, 90 per cent of cases. Thus, a significant false-negative rate exists with either primer which can be reduced by using both. We also found a difference in the efficiency of detection in different types of lymphoma; only 87 per cent of mucosa-associated lymphomas and centroblastic/centrocytic lymphomas were shown to be monoclonal, whereas all of the other lymphoma types tested were positive using one or both methods. We conclude that PCR detection of B-cell monoclonality allows rapid analysis of tissue samples, including paraffin-processed material. False-negative results which occur in some types of lymphoma can be reduced by the use of two or more primer combinations.
利用聚合酶链反应(PCR)检测B细胞单克隆性有望快速且经济高效地将单克隆性B细胞疾病与多克隆性B细胞疾病区分开来。然而,该方法的效率尚未得到充分评估,尤其是在疾病类型和PCR引物选择方面。我们基于使用框架2(Fr2)和框架3(Fr3)区域引物扩增免疫球蛋白重链基因评估了两种方法。对94例低度B细胞淋巴瘤的冷冻组织样本进行了研究,所有样本先前经Southern印迹分析已显示为单克隆性。使用Fr2引物,我们能够在85%的病例中显示单克隆性;使用Fr3引物,为80%;在单独反应中同时使用这两种技术,则为90%。因此,单独使用任何一种引物都存在显著的假阴性率,而同时使用两种引物可降低该比率。我们还发现不同类型淋巴瘤的检测效率存在差异;仅87%的黏膜相关淋巴瘤和中心母细胞/中心细胞淋巴瘤显示为单克隆性,而使用一种或两种方法检测的所有其他淋巴瘤类型均为阳性。我们得出结论,PCR检测B细胞单克隆性可对组织样本(包括石蜡处理的材料)进行快速分析。通过使用两种或更多引物组合,可减少某些类型淋巴瘤中出现的假阴性结果。