多聚酶链反应检测淋巴细胞白血病微小残留分析

Analysis of the Minimal Residuals of ALL Delected by PCR

根据IgH、TCR克隆性基因重排原理,用半重叠PCR和单轮PCR技术分别对同一份ALL患儿的骨髓标本进行检测(共25例),并对两法的检出率进行了比较。结果:半重叠PCR阳性捡出率为80％,单轮PCR阳性检出率为52％,前者较后者提高了28％,尤其对ALL-MRD组的检测,半重叠PCR的阳性检出率有显著提高。结果表明,半重叠PCR双轮扩增IgH、TCRγ基因重排对于MRD的检测起到增敏效果,可减少非特异性产物的生成,大大提高了MRD的阳性检出率,并与单轮PCR同样具有简便、快速以及用DNA量少,对DNA模板质量要求低,无放射性污染等优点。对区别肿瘤细胞的T、B细胞源性TCRβ基因重排尤其有价值。

In this study, the single-ture primers and semi-nested primers were used in the PCR procedure for detecting IgH and TCR gene rearrangements in the same DNA sample from bone marrow of 25 children suffering from acute lymphoblastic leukemia (ALL), and the positive rates were compared. Results, the positive detectable rate of semi-nested PCR was 80%,that of single-ture PCR was 52%.The former increased by 28% as com pared with the latter. Particularly, for the detecting of ALL-MRD group, the positive detectable rate of the semi-nested PCR remarkably improved. The results showed that semi-nested PCR is more sensitive and specific in detecting IgH and TCR gene rearrangements of ALL-MRD. But for distinguishing T、B cellular lineage of monoclonal cells,TCR βgene rearrangement was more significant.