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100例成人急性白血病中重链免疫球蛋白基因(IgH)第三互补决定区(CDR-III)的扩增

Amplification of third-complementary-determining-region (CDR-III) of heavy chain immunoglobulin gene (IgH) in one hundred adult acute leukemias.

作者信息

Martinelli G, Testoni N, Farabegoli P, Zaccaria A, Amabile M, Visani G, Pelliconi S, Raspadori D, Manfroi S, Vittone A, Terragna C, Tura S

机构信息

Institute of Hematology L. e A. Seràgnoli, Ospedale S. Orsola University of Bologna, Italy.

出版信息

Leuk Lymphoma. 1997 Jun;26(1-2):131-9. doi: 10.3109/10428199709109168.

Abstract

We applied a simple polymerase chain reaction (PCR) based method for detecting immunoglobulin heavy-chain (IgH) gene rearrangement, using its CDR-III region to assess B-cell clonality in a series of 100 acute lymphoblastic leukemias (ALL) (84 B-cell lineage, 4 null-ALL and 12 T-ALL). The amplified CDR-III regions can be generated in all the B-lineage ALL and separated by size by polyacrylamide gel electrophoresis (PAGE), thereby providing a specific diagnostic marker for each B cell clone. Size heterogeneity resulting from independent IgH rearrangement events and the high resolution power after electrophoresis and silver staining of the PAGE gels can be used to generate a "fingerprint" of the PCR fragments representing either the spectrum of B-cell clonality in complex populations of B lymphocytes or the partially genomic configuration of the VH-N-DH-N-JH region. At diagnosis, we found the presence of one clonal IgH heavy-chain gene rearrangement in 80 B-cell lineage and null ALL and a biclonal rearrangement in 8 cases. The CDR-III bands were of sizes ranging from 80 to 130 bp. The PCR analysis of the IgH gene enabled us to obtain a CDR-III leukemia specific product in all cases, thereby providing a specific and diagnostic marker for each B-cell clone.

摘要

我们应用了一种基于简单聚合酶链反应(PCR)的方法来检测免疫球蛋白重链(IgH)基因重排,利用其互补决定区III(CDR-III)区域评估100例急性淋巴细胞白血病(ALL)(84例B细胞系、4例无丙种球蛋白血症ALL和12例T细胞ALL)中的B细胞克隆性。在所有B细胞系ALL中均可产生扩增的CDR-III区域,并通过聚丙烯酰胺凝胶电泳(PAGE)按大小分离,从而为每个B细胞克隆提供特异性诊断标志物。由独立的IgH重排事件导致的大小异质性以及PAGE凝胶电泳和银染后的高分辨率,可用于生成代表B淋巴细胞复杂群体中B细胞克隆性谱或VH-N-DH-N-JH区域部分基因组构型的PCR片段“指纹”。在诊断时,我们发现80例B细胞系和无丙种球蛋白血症ALL中存在一种克隆性IgH重链基因重排,8例存在双克隆重排。CDR-III条带大小在80至130 bp之间。对IgH基因的PCR分析使我们能够在所有病例中获得CDR-III白血病特异性产物,从而为每个B细胞克隆提供特异性诊断标志物。

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