Stetlet-Stevenson M, Raffeld M, Cohen P, Cossman J
Laboratory of Pathology, National Cancer Institute, Bethesda, MD 20892.
Blood. 1988 Nov;72(5):1822-5.
To detect occult lymphoma, the polymerase chain reaction (PCR) technique was used to amplify joined bcl-2/JH DNA sequences at the juncture of the t(14:18) translocation in follicular lymphoma. Using the heat-stable DNA polymerase Taq and automated cycling of the reaction, we were able to detect as few as one to two copies of bcl-2/JH. Under these conditions, PCR proved to be at least 10,000-fold more sensitive than either conventional flow cytometry or Southern blot restriction analysis. In addition, genomic DNA sequences of four lymphomas confirmed that the size of the amplified segment serves as a tumor marker. Direct application of PCR to patient staging revealed occult malignant lymphoma in tissue otherwise considered uninvolved by standard criteria. We conclude that the striking enhancement in diagnostic sensitivity attained by DNA amplification can serve as a valuable adjunct to the staging and clinical monitoring of follicular lymphoma.
为检测隐匿性淋巴瘤，采用聚合酶链反应（PCR）技术扩增滤泡性淋巴瘤中t(14:18)易位连接处的连接bcl-2/JH DNA序列。使用热稳定DNA聚合酶Taq并对反应进行自动循环，我们能够检测到低至一到两个bcl-2/JH拷贝。在这些条件下，PCR被证明比传统流式细胞术或Southern印迹限制性分析至少敏感10000倍。此外，四个淋巴瘤的基因组DNA序列证实扩增片段的大小可作为肿瘤标志物。将PCR直接应用于患者分期，发现在标准标准认为未受累的组织中存在隐匿性恶性淋巴瘤。我们得出结论，DNA扩增所实现的诊断敏感性的显著提高可作为滤泡性淋巴瘤分期和临床监测的有价值辅助手段。
