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[B细胞克隆性的聚合酶链反应定义:哪种电泳方法更佳]

[PCR definition of B-cell clonality: which electrophoretic method is better].

作者信息

Sidorova Iu V, Nikitin E A, Ryzhikova N V, Sudarikov A B

出版信息

Ter Arkh. 2008;80(7):43-7.

Abstract

AIM

To compare diagnostic efficacy of B-cell clonality determination in application of different electrophoretic methods.

MATERIAL AND METHODS

B-cell clonality was determined with different techniques in 89 patients having B-cell lymphoma (n = 48), B-cell lymphoma in remission (n = 11), reactive processes (n = 24), lymphogranulomatosis and T-cell lymphoma (n = 6). Healthy donors served control. Clonality was defined by rearrangements of Ig heavy chain genes with usage of primers to FR2 region of Ig gene. Electrophoretic methods were the following: agarose electrophoresis and SSCP.

RESULTS

Clonality was detected in the presence of more than 5.9% clonal cells from total count of mononuclear cells or more than 20% of clonal cells from total number of B-lymphocytes in the sample. We achieved detectability of B-clonality in B-cell tumors 87.5% (42 of 48 cases). False negative tests were also investigated with kits of primers to FR1 and FR3 regions, but clonality was not determined. Among reactive processes, non-B-cell tumors and healthy donors clonality was found in 1 of 41 cases. This patient had acute respiratory viral infection. Agarose electrophoresis and SSCP test results coincided.

CONCLUSION

Determination of B-cell clonality with agarose electrophoresis proved to be a simple, reliable, cost-effective and reproducible method of differential diagnosis of tumor and reactive lymphoproliferation. This method is not suitable for verification of tumor remission or assessment of minimal residual disease because of its low sensitivity.

摘要

目的

比较不同电泳方法在B细胞克隆性检测中的诊断效能。

材料与方法

采用不同技术对89例患者进行B细胞克隆性检测,其中B细胞淋巴瘤患者48例,缓解期B细胞淋巴瘤患者11例,反应性病变患者24例,淋巴肉芽肿病和T细胞淋巴瘤患者6例。健康供者作为对照。通过使用针对Ig基因FR2区域的引物检测Ig重链基因重排来确定克隆性。电泳方法如下:琼脂糖电泳和单链构象多态性分析(SSCP)。

结果

当样本中克隆细胞占单核细胞总数的比例超过5.9%或占B淋巴细胞总数的比例超过20%时,可检测到克隆性。在B细胞肿瘤中,我们检测到B细胞克隆性的比例为87.5%(48例中的42例)。还使用针对FR1和FR3区域的引物试剂盒研究了假阴性检测,但未确定克隆性。在反应性病变、非B细胞肿瘤和健康供者中,41例中有1例检测到克隆性。该患者患有急性呼吸道病毒感染。琼脂糖电泳和SSCP检测结果一致。

结论

琼脂糖电泳检测B细胞克隆性是一种简单、可靠、经济高效且可重复的肿瘤与反应性淋巴细胞增殖鉴别诊断方法。由于其灵敏度较低,该方法不适用于验证肿瘤缓解或评估微小残留病。

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