Routine application of polymerase chain reaction in the diagnosis of monoclonality of B-cell lymphoid proliferations.
Routine application of polymerase chain reaction in the diagnosis of monoclonality of B-cell lymphoid proliferations.
复制标题
聚合酶链反应在 B 细胞淋巴增殖单克隆性诊断中的常规应用。
DOI：
10.1097/00019606-199503000-00005
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
F. Menestrina
中科院分区：
文献类型：
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作者：
A. Achille;A. Scarpa;M. Montresor;M. Scardoni;G. Zamboni;M. Chilosi;P. Capelli;G. Franzin;F. Menestrina
我们评估了四种聚合酶链式反应(PCR)方法在检测冰冻和石蜡包埋的B细胞淋巴增殖中的单克隆性的效率。这些方法(称为FR3、FR3A、FR2和FR1)是基于重排的免疫球蛋白重链基因的扩增，使用识别框架区域I、II或III的引物。FR3、FR3A和FR2方法分别在51%、72%和67%的冷冻淋巴瘤DNA中重复检测到单克隆性。未观察到假阳性。FR2和FR3A相结合的方法将这一数字提高到85%。石蜡包埋淋巴瘤也得到了类似的结果。FR1法的重复性不理想。所有的聚合酶链式反应方法的效率因淋巴瘤类型而异。小/中细胞性淋巴瘤的检出率最高，中心型滤泡性淋巴瘤的检出率最低。限制性稀释法检测到占非淋巴细胞性B细胞DNA的5%，占多克隆B细胞DNA的20%。一种诊断方案可能包括快速且经济有效的聚合酶链式反应筛查，特别是在细针抽吸物或内窥镜活检中观察到的不明小细胞淋巴样增殖的情况下。这也将减少患者要求获得非固定活检的次数。
We evaluated four polymerase chain reaction (PCR) methods for their efficiency in detecting monoclonality in a well-characterized panel of frozen and paraffin-embedded B-cell lymphoid proliferations. These approaches (referred to as FR3, FR3A, FR2, and FR1) are based on amplification of rearranged immunoglobulin heavy chain genes, using primers recognizing framework regions I, II, or III. FR3, FR3A and FR2 approaches reproducibly detected monoclonality in 51%, 72%, and 67% of DNAs from frozen lymphomas, respectively. No false-positives were observed. The combination of FR2 and FR3A methods raised the figure to 85%. Comparable results were obtained using paraffin-embedded lymphomas. Reproducibility of FR1 approach was unsatisfactory. The efficiency of all PCR approaches varied depending on lymphoma type. The highest detection rate was in small/intermediate cell and the lowest in centro-follicular lymphomas. Limiting dilution assays showed that PCR methods were able to detect monoclonal B-cell DNA representing 5% of nonlymphoid and 20% of polyclonal B-cell DNA. A diagnostic protocol may include quick and cost-effective PCR screening, particularly in cases of undetermined small cell lymphoid proliferations observed in fine needle aspirates or endoscopic biopsies. This would also reduce call-up of patients to obtain unfixed biopsies.