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通过聚合酶链反应检测石蜡包埋组织中的克隆性B细胞群体。

Detection of clonal B cell populations in paraffin-embedded tissues by polymerase chain reaction.

作者信息

Küppers R, Zhao M, Rajewsky K, Hansmann M L

机构信息

I. Medizinische Universitätsklinik Köln, Germany.

出版信息

Am J Pathol. 1993 Jul;143(1):230-9.

Abstract

A method was established to detect clonal B cell populations in frozen and paraffin-embedded tissues. The method is based on the polymerase chain reaction amplification of rearranged VH and V kappa genes using V gene family-specific primers. Monoclonal B cell populations could be detected in peripheral blood lymphocyte DNA in all 16 cases of B-CLL and immunocytoma investigated and in 8 of 10 cases of B cell non-Hodgkin's lymphoma using frozen and paraffin sections. The amplification of V kappa rearrangements in addition to the VH amplification is a useful tool to verify the results of the heavy chain rearrangement and to detect proliferation of a B cell clone in cases in which no VH product was obtained. In spite of the degradation of DNA in paraffin-embedded tissues, we were able to find amplified polymerase chain reaction products of about 350 bp length in 8 of 10 cases analyzed. The method presented here may be helpful in routine diagnosis of B cell non-Hodgkin's lymphoma using frozen or paraffin-embedded specimens.

摘要

建立了一种检测冷冻组织和石蜡包埋组织中克隆性B细胞群体的方法。该方法基于使用V基因家族特异性引物对重排的VH和Vκ基因进行聚合酶链反应扩增。在所研究的16例B细胞慢性淋巴细胞白血病(B-CLL)和免疫细胞瘤以及10例B细胞非霍奇金淋巴瘤中的8例中,利用冷冻切片和石蜡切片,可在外周血淋巴细胞DNA中检测到单克隆B细胞群体。除了VH扩增外,Vκ重排的扩增是验证重链重排结果以及在未获得VH产物的情况下检测B细胞克隆增殖的有用工具。尽管石蜡包埋组织中的DNA发生了降解,但在分析的10例病例中的8例中,我们仍能够找到长度约为350 bp的扩增聚合酶链反应产物。本文介绍的方法可能有助于使用冷冻或石蜡包埋标本对B细胞非霍奇金淋巴瘤进行常规诊断。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b351/1886961/685cfc7861ff/amjpathol00067-0241-a.jpg

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