Segal G H, Scott M, Jorgensen T, Braylan R C
Department of Pathology and Laboratory Medicine, University of Florida College of Medicine, Gainesville.
Diagn Mol Pathol. 1994 Mar;3(1):15-21. doi: 10.1097/00019606-199403010-00004.
We observed a potentially misinterpretable polymerase chain reaction (PCR) amplification product generated with standard primers used to detect the major breakpoint region (mbr) of chromosomal translocation t(14;18). This unexpected phenomenon was initially detected during attempts to transform follicular lymphomas in vitro with Epstein-Barr virus (EBV). Additional studies were performed using the EBV-producing cell line MCUV5, cell lines from EBV-transformed normal B-lymphocytes, and an excised lymph node from a patient with documented EBV-associated infectious mononucleosis. These samples consistently produced a 167-base pair product, which was indistinguishable from a t(14;18) lymphoma product when viewed on ethidium bromide-stained gels. Through DNA sequencing and gene bank analysis, the product was identified as a portion of the EBV genome. A mbr-specific 20-base oligonucleotide probe was able to discriminate between true translocations and the EBV-related amplifications. These results underscore the importance of employing a specific detection system, and comprehensively screening primers when working with PCR.
我们观察到，在用用于检测染色体易位t(14;18)主要断裂点区域（mbr）的标准引物进行聚合酶链反应（PCR）时，产生了一种可能会被误判的扩增产物。这种意外现象最初是在尝试用爱泼斯坦-巴尔病毒（EBV）体外转化滤泡性淋巴瘤的过程中被检测到的。我们使用产生EBV的细胞系MCUV5、来自EBV转化的正常B淋巴细胞的细胞系以及一名有记录的EBV相关传染性单核细胞增多症患者的切除淋巴结进行了进一步研究。这些样本始终产生一种167个碱基对的产物，在溴化乙锭染色的凝胶上观察时，它与t(14;18)淋巴瘤产物无法区分。通过DNA测序和基因库分析，该产物被鉴定为EBV基因组的一部分。一种mbr特异性的20碱基寡核苷酸探针能够区分真正的易位和与EBV相关的扩增。这些结果强调了在进行PCR时采用特定检测系统以及全面筛选引物的重要性。
