实时荧光定量PCR检测急性淋巴细胞白血病患者TCR VγI-Jγ基因重排

目的　探索提高急性淋巴细胞白血病 (ALL)微量残留病 (MRD)检测技术。方法　构建实时荧光定量聚合酶链反应 (PCR)检测TCRVγI Jγ基因重排技术并定量分析 36例ALL患者的检测结果。结果　建立的实时荧光定量PCR方法的灵敏度为 10 -4水平。 36例患者荧光定量PCR方法检测结果初治组为 (7.38± 6 .6 5 )× 10 -2 ,完全缓解 (CR)组为 (1.0 2± 1.0 8)× 10 -2 ,造血干细胞移植(HSCT)组为 (3.89± 5 .6 5 )× 10 -3 。CR组和HSCT组TCRVγI Jγ基因重排水平显著低于初治组 (P值均为 0 .0 0 1) ,HSCT组MRD水平显著低于CR组 (P <0 .0 5 )。 6例HSCT后检测阳性病例中 2例MRD水平 <1× 10 -3 ,获长期无病生存 ,另 4例MRD水平较高患者一年内均出现复发。结论　所建立的实时荧光定量PCR方法简便、快速、灵敏及特异 ;实时荧光定量检测缓解期ALL患者MRD水平对预测预后有一定的意义

Establishment of a real time quantitative-PCR assay for detection of TCR VγI-Jγ gene rearrangement in acute lymphoblastic leukemia patients

OBJECTIVE: To improve the techniques for minimal residual disease (MRD) detection in acute lymphoblastic leukemia (ALL). METHODS: A real time quantitative PCR method was established for quantifying the clonal TCRVgammaI-Jgamma gene rearrangement in 36 ALL patients. RESULTS: The sensitivity of the established real time quantitative PCR was at 10(-4) level. The amount of TCRVgammaI-Jgamma gene rearrangement in newly diagnosed group, complete remission (CR) group and post hematopoietic stem cell transplantation (HSCT) group was (7.38 +/- 6.65) x 10(-2), (1.02 +/- 1.08) x 10(-2) and (3.89 +/- 5.65) x 10(-3) level, respectively. and the amount in newly diagnosed group was higher than that in CR group and HSCT group (P = 0.001). The MRD level of ALL patients in CR group was higher than that in HSCT group (P = 0.022). MRD can be detected in 6 ALL patients after HSCT, 2 of them with low MRD level (< 1 x 10(-3)) survived long disease-free survival, the other 4 with high MRD level relapsed within one year. CONCLUSION: The established real time quantitative PCR assay is simple, rapid, sensitive and specific. Use of this assay to evaluate MRD in the remission ALL cases is helpful for prognosis prediction.