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一种基于聚合酶链反应的快速技术,用于检测αβ和γδ两种类型皮肤T细胞淋巴瘤中的克隆性T细胞受体基因重排。

A rapid polymerase chain reaction-based technique for detecting clonal T-cell receptor gene rearrangements in cutaneous T-cell lymphomas of both the alpha beta and gamma delta varieties.

作者信息

Yu R C, Alaibac M

机构信息

Unit of Dermatology, Royal Postgraduate Medical School, London, England.

出版信息

Diagn Mol Pathol. 1996 Jun;5(2):121-6. doi: 10.1097/00019606-199606000-00007.

Abstract

T-cell receptor-gamma gene rearrangements provide specific clonal markers for a variety of lymphoid malignancies. T-cell receptor gene rearrangements in patients with cutaneous T-cell lymphoma were examined using conventional Southern blot analysis and a newly developed polymerase chain reaction (PCR)-based technique. The oligoprimers amplified a rearranged V gamma and J gamma segment (including the N region) of the T-cell receptor-gamma gene, and products were resolved using high-resolution nondenaturing polyacrylamide gel electrophoresis. Our results demonstrated concordance between the two techniques in 10 patients with cutaneous T-cell lymphomas (including nine cases of C beta and one case of delta 2 TCR gene rearrangements) and 10 negative controls. In the present study, we have shown that this PCR-based method provides a highly sensitive, specific technique for the detection of T-cell clones of both the alpha beta and gamma delta varieties and could be used in both fresh and formalin-fixed, paraffin-embedded tissues. It is estimated that this PCR-based technique is 10 to 50 times more sensitive than conventional Southern blot analysis in the detection of small T-cell clones.

摘要

T细胞受体γ基因重排为多种淋巴系统恶性肿瘤提供了特异性的克隆标记。我们运用传统的Southern印迹分析法和一种新开发的基于聚合酶链反应(PCR)的技术,检测了皮肤T细胞淋巴瘤患者的T细胞受体基因重排情况。寡核苷酸引物扩增了T细胞受体γ基因的重排Vγ和Jγ片段(包括N区),产物通过高分辨率非变性聚丙烯酰胺凝胶电泳进行分离。我们的结果表明,在10例皮肤T细胞淋巴瘤患者(包括9例Cβ和1例δ2 TCR基因重排)以及10例阴性对照中,两种技术的检测结果一致。在本研究中,我们已经证明,这种基于PCR的方法为检测αβ和γδ两种类型的T细胞克隆提供了一种高度灵敏、特异的技术,并且可用于新鲜组织以及福尔马林固定、石蜡包埋的组织。据估计,在检测小T细胞克隆方面,这种基于PCR的技术比传统的Southern印迹分析法灵敏10至50倍。

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