多聚酶链反应和限制酶分析急性白血病T细胞受体γ链基因重排

ANALYSIS OF T CELL RECEPTOR γ CHAIN GENE REARRANGEMENT USING POLYMERASE CHAIN REACTION AND RESTRICTION ENZYMES DIGEST IN ACUTE LEUKEMIA

选用TCR γ基因保守区设计的引物,用PCR扩增30例白血病患者的T细胞受体(TCR)γV-N-J重排区。再用5种限制性内切酶消化。分析白血病参加重排的V区片段,同时观察重排基因与形态和免疫分型的关系。结果表明,简单的1对引物和5种限制酶就可以确定TCRγV区大部分片段的类型。此法检测时间短,获得结果迅速,敏感性强。重排的V区类型可以作为基因标志,以研究白血病治疗过程中的微小残留细胞和克隆进化。

We used a pair of consensus primers to amplify rearrangements involving the T cell receptor γ chain (TCRγ) V-N-J genes from 30 cases of acute leukemia. The PCR products were digested with five restriction enzymes and analyzed by electropherosis. The relationship between the gene rearrangement and classification of morphology and immunology in the leukemias were investigated. The results demonstrate that it is possible to determine V gene quickly and sensitively by restriction digestion of PCR products. This method can be used to study the leukemia residual cell, a molecular marker during the course of treatment, and the leukemia clonal evolution.