半重叠式多重引物PCR和基因扫描技术在T细胞淋巴瘤诊断中的应用

Detection of T cell receptor γ gene rearrangement in paraffin-embedded tissues of lymphoma by semi-nested multicolor multiplex PCR and Gene-Scan

[目的]探讨半重叠式多重荧光多重引物PCR和基因扫描技术在T细胞淋巴瘤诊断中的应用.[方法]从普通石蜡切片组织标本中提取基因组DNA,用半重叠式多重荧光多重引物PCR扩增,利用DNA测序仪对其产物进行基因扫描分析.[结果]37例T细胞淋巴瘤中27例检测出有克隆性重排;25例B细胞淋巴瘤中2例呈阳性;10例何杰金氏淋巴瘤和7例反应性增生病例均呈阴性.[结论]在T细胞受体γ基因重排克隆性检测中,半重叠式多重荧光多重引物PCR及基因扫描技术具有样本来源便利,灵敏度高,特异性强,耗时短等优点,可作为T细胞淋巴瘤的诊断及鉴别诊断的辅助检测手段.

OBJECTIVE To explore the application of semi-nested multicolor multiplex PCR and Gene-Scan in the detection of T cell receptor γ （TCR-γ） rearrangement. METHODS DNA from paraffin-embedded tissues were extracted, and amplified by semi-nested multicolor multiplex PCR. The fluorescence labeled PCR products were separated with an automated sequencing system, and analyzed by Gene-Scan. RESULTS 27 out of 37 cases of T-cell lymphoma and 2 out of 25 cases of B-cell lymphoma were positive for clonal TCR-γ rearrangement, 10 cases of Hodgkin lymphoma and 7 cases of reactive lymphoid hyperplasia were negative. These results were corresponding to the detection by southern blot. CONCLUSION Semi-nested multicolor multiplex PCR-Gene-Scan technique is a sensitive and convenient tool for TCR-γ rearrangement analysis, and can be useful in diagnosis and differential diagnosis of T cell lymphoma.