Segal G H, Shick H E, Tubbs R R, Fishleder A J, Stoler M H
Department of Pathology, Cleveland Clinic Foundation, Ohio.
Diagn Mol Pathol. 1994 Sep;3(3):170-7.
Determination of clonality in B-cell lymphomas is a useful diagnostic adjunct. In situ hybridization (ISH) for the detection of kappa and lambda mRNAs has the potential to overcome some common specimen-related limitations in clonal assessment. Tritium-labeled antisense cRNA probes directed at conserved segments of the constant regions of the kappa and lambda mRNAs were used in an autoradiographic method to detect B-cell clonality. Using these probes, we analyzed 103 formalin-fixed, paraffin-embedded biopsy samples, and the results were subsequently compared to available immunophenotypic (all cases) and genotypic (50 cases) data. Of 103 samples, 82 (80%) had adequate RNA preservation as determined by actin RNA signals, and 73 (89%) of the 82 cases demonstrated concordant clonality assignment by both ISH and immunophenotyping. The remaining nine cases showed a specific form of discordance in that each exhibited no protein (Ig) expression but had evidence of mRNA immunoglobulin light-chain expression. Forty-five (90%) of 50 cases evaluated for immunoglobulin and T-cell receptor beta-gene rearrangements demonstrated concordant results with respect to clonality assignment by ISH. Thus, ISH demonstrates adequate sensitivity with respect to traditional methods of clonality assessment. However, its practical utility awaits the development of nonradioactive detection methods with adequate sensitivity to improve turnaround time.
确定B细胞淋巴瘤的克隆性是一种有用的辅助诊断方法。用于检测kappa和lambda mRNA的原位杂交（ISH）有潜力克服克隆性评估中一些常见的与标本相关的局限性。针对kappa和lambda mRNA恒定区保守片段的氚标记反义cRNA探针被用于放射自显影方法来检测B细胞克隆性。使用这些探针，我们分析了103份福尔马林固定、石蜡包埋的活检样本，随后将结果与现有的免疫表型（所有病例）和基因分型（50例）数据进行比较。在103份样本中，根据肌动蛋白RNA信号确定，82份（80%）有足够的RNA保存，82例中的73例（89%）通过ISH和免疫表型分析显示出一致的克隆性判定。其余9例表现出一种特殊形式的不一致，即每例均无蛋白（Ig）表达，但有mRNA免疫球蛋白轻链表达的证据。在评估免疫球蛋白和T细胞受体β基因重排的50例中，45例（90%）在克隆性判定方面通过ISH显示出一致的结果。因此，ISH相对于传统的克隆性评估方法显示出足够的敏感性。然而，其实际应用有待于开发出具有足够敏感性以缩短周转时间的非放射性检测方法。
