Luthra R, Hai S, Pugh W C
Department of Pathology, University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Diagn Mol Pathol. 1995 Mar;4(1):4-7. doi: 10.1097/00019606-199503000-00003.
The utility of polymerase-mediated assays in the detection of the t(11;14) involving the bcl-1 major translocation cluster (bcl-1 MTC) was evaluated by analyzing DNA from 33 patients with mantle cell lymphoma, 14 patients with other non-Hodgkin's lymphomas, and five patients with reactive lymphoid hyperplasia. The polymerase chain reaction (PCR) assay was performed using a consensus immunoglobin heavy-chain joining region primer in conjunction with a chromosome 11 specific oligonucleotide primer flanking the translocation site. The sensitivity and specificity of the assay were confirmed by correlation of the (PCR) assay data with restriction analysis. Rearrangements at the bcl-1 MTC were detected in 13 (39%) of 33 cases of mantle cell lymphoma by PCR and in 13 (48%) of 27 cases by restriction analysis. Amplicons were detectable by PCR in 85% (11 of 13) of the cases shown to be bcl-1 rearranged by restriction analysis. Failure to detect amplification products in DNA samples from non-mantle cell lymphomas and reactive follicular hyperplasia further confirmed the specificity of the assay. Sequential hybridization of the PCR products with oligonucleotide probes 3' to the bcl-1 MTC primer revealed that the breakpoints in the bcl-1 MTC were clustered around an Sst I restriction site over a range of 170 base pairs. The study demonstrates that PCR-mediated assay for the detection of the t(11;14) at the bcl-1 MTC is specific and sensitive and can be used as an adjunct to restriction analysis in routine diagnostics.
通过分析33例套细胞淋巴瘤患者、14例其他非霍奇金淋巴瘤患者及5例反应性淋巴组织增生患者的DNA,评估了聚合酶介导的检测法在检测涉及bcl-1主要易位簇(bcl-1 MTC)的t(11;14)中的效用。使用共有免疫球蛋白重链连接区引物结合位于易位位点侧翼的11号染色体特异性寡核苷酸引物进行聚合酶链反应(PCR)检测。通过将PCR检测数据与限制性分析结果相关联,证实了该检测法的敏感性和特异性。在33例套细胞淋巴瘤中,有13例(39%)通过PCR检测到bcl-1 MTC重排,27例中有13例(48%)通过限制性分析检测到重排。在经限制性分析显示bcl-1重排的病例中,85%(13例中的11例)通过PCR可检测到扩增产物。在非套细胞淋巴瘤和反应性滤泡增生的DNA样本中未能检测到扩增产物,进一步证实了该检测法的特异性。将PCR产物与bcl-1 MTC引物3'端的寡核苷酸探针进行顺序杂交,结果显示bcl-1 MTC中的断点聚集在一个170个碱基对范围内的Sst I限制性位点周围。该研究表明,用于检测bcl-1 MTC处t(11;14)的PCR介导检测法具有特异性和敏感性,可在常规诊断中作为限制性分析的辅助方法。