Wan J H, Trainor K J, Brisco M J, Morley A A
Department of Haematology, Flinders Medical Centre, Bedford Park, South Australia.
J Clin Pathol. 1990 Nov;43(11):888-90. doi: 10.1136/jcp.43.11.888.
The polymerase chain reaction (PCR) was used to develop a simple technique for detecting monoclonality at the DNA level in B lymphocyte populations in formalin fixed, paraffin wax embedded material. Sections were dewaxed and dehydrated, and the DNA was extracted by boiling in water for 45 minutes. A semi-nested PCR was performed to amplify the V-D-J region of the immunoglobulin heavy chain gene. The product was electrophoresed and viewed under ultraviolet light after ethidium bromide staining. Specimens from 26 B cell lymphomas produced a monoclonal band in 24 cases and no amplification in two cases; monoclonality was specific for this disorder. Specimens from seven T cell lymphomas produced no amplification; specimens from nine reactive nodes produced a broad smear of polyclonal material; and specimens from 12 cases of carcinoma produced either no amplification or polyclonal material. As detection of monoclonality is strongly suggestive of neoplastic disease, this technique is likely to be of value in routine diagnosis, because of its speed, simplicity, and applicability to fixed, embedded material.
聚合酶链反应（PCR）被用于开发一种简单技术，以在福尔马林固定、石蜡包埋材料中的B淋巴细胞群体的DNA水平检测单克隆性。切片脱蜡并脱水，DNA通过在水中煮沸45分钟来提取。进行半巢式PCR以扩增免疫球蛋白重链基因的V-D-J区域。产物经电泳，并在溴化乙锭染色后在紫外线下观察。来自26例B细胞淋巴瘤的标本在24例中产生单克隆条带，2例无扩增；单克隆性是这种疾病的特异性表现。来自7例T细胞淋巴瘤的标本无扩增；来自9个反应性淋巴结的标本产生多克隆物质的宽条带；来自12例癌的标本要么无扩增要么产生多克隆物质。由于单克隆性的检测强烈提示肿瘤性疾病，这项技术因其速度、简便性以及对固定、包埋材料的适用性，在常规诊断中可能具有价值。
