Taube T, Seeger K, Beyermann B, Hanel C, Duda S, Linderkamp C, Henze G
Department of Pediatric Oncology/Hematology, Charité-Virchow Medical Center, Humboldt-University Berlin, Germany.
Leukemia. 1997 Nov;11(11):1978-82. doi: 10.1038/sj.leu.2400825.
A rapid and simple multiplex polymerase chain reaction (PCR) is described that is capable of identifying the six most frequent rearrangements of the T cell receptor (TCR)-delta gene segments in childhood acute lymphoblastic leukemia (ALL). The PCR products amplified in a single reaction are of different size for each TCR-delta gene rearrangement. Therefore, they are readily and unambiguously distinguished after agarose gel electrophoresis and assigned to a specific V-D-J gene rearrangement. There is no need for labor-intensive and time-consuming Southern blot hybridization or nested PCR. To evaluate the multiplex assay we chose 45 DNA samples of childhood ALL analyzed beforehand for TCR-delta gene rearrangements by Southern blot and single PCR of which 30 showed TCR-delta gene rearrangements. The multiplex PCR results corresponded to the Southern blot and single PCR analyses. The described multiplex PCR enables the detection of clonal markers in about 50% of patients in order to monitor minimal residual disease (MRD) in prospective studies with a high turnover of samples.
本文描述了一种快速简便的多重聚合酶链反应（PCR），它能够识别儿童急性淋巴细胞白血病（ALL）中T细胞受体（TCR）-δ基因片段的六种最常见重排。在单一反应中扩增的PCR产物对于每种TCR-δ基因重排具有不同大小。因此，它们在琼脂糖凝胶电泳后很容易且明确地被区分，并被指定为特定的V-D-J基因重排。无需进行劳动强度大且耗时的Southern印迹杂交或巢式PCR。为了评估多重检测方法，我们选择了45份儿童ALL的DNA样本，这些样本之前已通过Southern印迹和单重PCR对TCR-δ基因重排进行了分析，其中30份显示有TCR-δ基因重排。多重PCR结果与Southern印迹和单重PCR分析结果一致。所描述的多重PCR能够在约50%的患者中检测到克隆标志物，以便在样本周转量大的前瞻性研究中监测微小残留病（MRD）。
