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套细胞淋巴瘤。基于快速聚合酶链反应的形态学异质性实体的基因分型。

Mantle cell lymphoma. Rapid polymerase chain reaction-based genotyping of a morphologically heterogeneous entity.

作者信息

Segal G H, Maiese R L

机构信息

Department of Pathology, University of Utah Health Sciences Center, Salt Lake City 84132, USA.

出版信息

Arch Pathol Lab Med. 1996 Sep;120(9):835-41.

PMID:9140288
Abstract

BACKGROUND

Mantle cell lymphoma is heterogeneous at the morphologic level. Since this B-cell lymphoma may be confused with other entities, ancillary molecular testing may be necessary for definitive diagnosis. A polymerase chain reaction-based method, which is less complicated and more rapid than that generally available for the detection of immunoglobulin heavy chain (IgH) and bcl-1 gene rearrangements, would be helpful in this process.

METHODS

Thirty-one mantle cell lymphoma samples (frozen or ethanol-preserved) from 29 patients were studied with two separate polymerase chain reaction assays using an air thermocycler and a low-volume, capillary-tube format for rapid DNA amplification. The reverse primer, JH, was common to both assays. The forward primers were directed to the IgH framework III variable region (VH-FRIII) and the bcl-1 gene major translocation cluster. Agarose gels were used to evaluate amplicon. Additional product verification was also performed.

RESULTS

Immunoglobulin heavy chain and major translocation cluster bcl-1 gene rearrangements were detected in all 29 (100%) and in 12 (41%) of 29 mantle cell lymphoma samples, respectively. Each VH-FRIII/JH assay required 26 minutes to complete, whereas the major translocation cluster bcl-1/JH reaction required only 21 minutes. The seemingly low yield of bcl-1 gene rearrangements is not unexpected since this assay only detects major translocation cluster breakpoints.

CONCLUSIONS

Presented is an extremely rapid, nonisotopic polymerase chain reaction-based method that detects IgH and major translocation cluster bcl-1 gene rearrangements in mantle cell lymphoma. Each polymerase chain reaction amplification was complete in 26 minutes or less, required only a 10-microL reaction volume, and exhibited adequate and specific product yield. This approach permits superior turnaround time and is thus advantageous in the clinical setting.

摘要

背景

套细胞淋巴瘤在形态学水平上具有异质性。由于这种B细胞淋巴瘤可能与其他实体瘤混淆,因此辅助分子检测对于明确诊断可能是必要的。一种基于聚合酶链反应的方法,比通常用于检测免疫球蛋白重链(IgH)和bcl-1基因重排的方法更简单、更快速,将有助于这一过程。

方法

使用空气热循环仪和低体积毛细管形式对来自29例患者的31份套细胞淋巴瘤样本(冷冻或乙醇保存)进行两项独立的聚合酶链反应检测,以进行快速DNA扩增。两种检测方法的反向引物JH相同。正向引物分别针对IgH框架III可变区(VH-FRIII)和bcl-1基因主要易位簇。使用琼脂糖凝胶评估扩增子。还进行了额外的产物验证。

结果

在所有29份(100%)套细胞淋巴瘤样本中均检测到免疫球蛋白重链和主要易位簇bcl-1基因重排,分别在29份样本中的12份(41%)中检测到。每个VH-FRIII/JH检测需要26分钟完成,而主要易位簇bcl-1/JH反应仅需21分钟。bcl-1基因重排的产量看似较低并不意外,因为该检测仅检测主要易位簇断点。

结论

本文介绍了一种基于聚合酶链反应的极其快速、非同位素的方法,可检测套细胞淋巴瘤中的IgH和主要易位簇bcl-1基因重排。每次聚合酶链反应扩增在26分钟或更短时间内完成,仅需10微升反应体积,并表现出足够且特异的产物产量。这种方法具有出色的周转时间,因此在临床环境中具有优势。

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