Dubus P, Labouyrie E, Bilhou-Nabera C, Belleannee G, Vergier B, Bernard P, de Mascarel A, Merlio J P
Service d'Anatomie Pathologique, CHU de Bordeaux, Pessac.
Ann Pathol. 1994;14(4):227-33.
Family-specific primers were used in polymerase chain reaction (PCR) to analyse clonality of immunoglobulin heavy chain (IgH) gene rearrangement. DNA templates were extracted from formalin-fixed paraffin-embedded biopsies from B-cell lymphomas (n = 19), T-cell lymphomas (n = 3), reactive lymphadenopathies (n = 20) and negative controls (n = 2). PCR was also performed on DNA extracted from fragments of the same biopsies that were either fixed in Bouin's solution (n = 24) or snap-frozen (n = 22). The latter were also studied for IgH gene rearrangement by the Southern blot technique. Polyclonal or clonal fragments were obtained from all frozen biopsies. No amplification was observed in 3 out of 44 formalin fixed specimens and 18 out of 24 Bouin fixed specimens. A clonal amplification was only observed in 16 out of 19 formalin-fixed B-cell lymphoma specimens. Therefore this technique allows to study B-cell clonality from formalin-fixed, embedded material.
使用家族特异性引物通过聚合酶链反应（PCR）分析免疫球蛋白重链（IgH）基因重排的克隆性。从B细胞淋巴瘤（n = 19）、T细胞淋巴瘤（n = 3）、反应性淋巴结病（n = 20）的福尔马林固定石蜡包埋活检组织以及阴性对照（n = 2）中提取DNA模板。还对用Bouin溶液固定（n = 24）或速冻（n = 22）的相同活检组织片段提取的DNA进行PCR。后者也通过Southern印迹技术研究IgH基因重排。从所有冷冻活检组织中获得了多克隆或克隆片段。在44份福尔马林固定标本中有3份未观察到扩增，在24份Bouin固定标本中有18份未观察到扩增。仅在19份福尔马林固定的B细胞淋巴瘤标本中的16份中观察到克隆扩增。因此，该技术可用于研究福尔马林固定、包埋材料中的B细胞克隆性。
