竞争性定量PCR检测TCRVγI-Jγ基因重排方法的建立及应用

Construction of a quantitative-PCR for detection of TCRV γI-Jγ gene rearrangement and its clinical application

目的为提高急性淋巴细胞白血病（ALL）微小残留检测的临床意义。方法建立竞争性定量聚合酶链反应（PCR）检测TCRVγI－Jγ基因重排技术并定量分析16例ALL病人残留白血病细胞。结果建立定量PCR方法的灵敏度为10-5水平。16例ALL缓解期残留白血病细胞为（427±364）×10-5病人。3例病人可检测出寡／亚克隆重排，白血病细胞定量为117×10-5、196×10-5及211×10-5水平，其中1例寡／亚克隆水平在病程中不断增加并导致3月后复发伴克隆演化。结论所建立定量PCR技术简便、快速、灵敏；定量检测ALL缓解期残留白血病有利于预后预测并可应用于寡／亚克隆的定量检测。

Objective To improve the clinical significance of detecting minimal residual disease (MRD) in acute lymphoblasticleukemia (ALL). Methods A competitive PCR method was constructed for quantitative clonal rearrangement of TCRV YI-J Y genein 16 ALL cases. Result The sensitivity of the established quantitative-PCR was at 10-5 level. In remission, the leukemic cells in 16ALL cases were (427±364)×10-5. The recurrence rate was higher in patients with leukemic cells M520 X 109/L than in those wilhleukemic cells <520 × 109/L. Oligoclonal/subclonal rearrangement can be detected in 3 cases whose leukemic cells were 117×10-5.196 × 10-5 and 211 × 10-5, respectively,in one of whom increasing of oligoclonal subclonal in the course of disease led to relapse 3months later clonal evolution. Conclusion The established quantitative PCR assay is simple, lepaid and sensitive. The quantitativeevaluation of ALL cases during the remission period may help for prediction of prognosis and also can be used for quantitative assayof the oligoclones /subclones.