琼脂糖凝胶电泳及SSCP法在淋巴瘤克隆性TCRγ基因重排检测中的应用

Application of agarose gel electrophoresis and SSCP in lymphoma clonal TCRγ gene rearrangement detecting

目的比较琼脂糖凝胶电泳和单链构象多态性分析(SSCP)法对淋巴瘤克隆性T细胞受体(TCR)γ基因重排的检测结果,探讨TCRγ基因重排的有效检测方法。方法从T、B细胞淋巴瘤及反应增生性淋巴组织中提取DNA,分别用两组TCRγ基因重排通用引物进行PCR扩增,扩增产物分别进行琼脂糖凝胶电泳和SSCP。结果T、B细胞淋巴瘤均出现克隆性TCRγ基因重排。SSCP法两组PCR产物在T细胞淋巴瘤中的阳性率分别为77.6%和75.9%,B细胞淋巴瘤均为10%;与SSCP相比,琼脂糖凝胶电泳中两组PCR扩增产物的假阳性率分别为15.3%和14.4%,所有反应增生性淋巴组织均出现假阳性。结论T、B细胞淋巴瘤中都存在克隆性TCRγ基因重排,仅用琼脂糖凝胶电泳检测可能出现假阳性,SSCP灵敏性较高。

Objective To compare the results of agarose gel electrophoresis and single stranded configuration pleomorphism（SSCP）analysis in detecting T cell receptor（TCR）γ gene rearrangement,explore the effective methods for TCRγ gene rearrangement detection.Method DNA extracted from T,B cell lymphomas and reactive lymphoid tissues were respectively submitted to two groups of TCRγgene rearrangement universal primers for PCR amplification,and PCR products were separately proceeded to agarose gel electrophoresis and SSCP.Results Clonal TCRγgene rearrangement presented both in T and B cell lymphomas.In SSCP of the two groups of PCR products,positive ratios of T cell lymphoma were 77.6% and 75.9%,respectively,while in B cell lymphomas,the positive ratios were both 10%.Compared to SSCP,false positive ratios in agarose gel electrophoresis were 5.3% and 14.4%,respectively.All the reactive lymphoid tissue emerged false positive.Conclusions Both T and B cell lymphomas exist clonal TCRγ gene rearrangement.Detection with agarose gel electrophoresis alone may result false positive,and the sensitivity of SSCP is higher.