筑巢式聚合酶链反应动态检测儿童微量残留白血病的研究

Dynamic detection of minimal residual disease in children with leukemia by using nested polymerase chain reaction

为探讨Ｔ细胞受体（ＴＣＲ）Ｖδ２－Ｄδ３基因重排检测对儿童急性淋巴细胞白血病（ＡＬＬ）分型诊断及微量残留病（ＭＲＤ）动态监测的意义，采用筑巢式聚合酶链反应（Ｎｅｓｔｅｄ－ＰＣＲ）技术，对４０例ＡＬＬ患儿进行了ＴＣＲＶδ２－Ｄδ３基因重排的检测，并对其中３０例在其完全缓解（ＣＲ）后进行了ＭＲＤ的动态监测。结果，４０例ＡＬＬ中３０例（７５％），２５例Ｂ－ＡＬＬ中２０例（８０％），４例Ｔ－ＡＬＬ中１例检出了克隆特异性ＴＣＲＶδ２－Ｄδ３基因重排。２７例初次ＴＣＲＶδ２－Ｄδ３基因重排检测阳性者中，５例ＣＲ后ＭＲＤ转阴，其中４例已终止化疗，观察至今已无病存活８５～１９９个月；８例ＭＲＤ转为弱阳性，其中３例曾骨髓复发，经治疗再次ＣＲ，已存活４１～１８７个月，仍在继续化疗和监测中；１３例ＭＲＤ持续阳性和１例ＭＲＤ由阴转阳者中７例（５０％）发生了骨髓复发并５例死亡。研究表明，ＡＬＬ患儿ＣＲ达３年以上，ＭＲＤ持续转阴者预后良好，并可以此作为终止化疗的可靠指标。若ＭＲＤ持续阳性或由阴转阳性时，提示有骨髓复发的可能性，应给予有效化疗。应用Ｎｅｓｔｅｄ－ＰＣＲ动态监测ＡＬＬ的ＭＲＤ，对预测复发、判断预后及指导化疗等有重要临床意义?

For exploring the relationship between the rearrangement of T cell recepter (TCR) Vδ 2 Dδ 3 in children with acute lymphoblastic leukemia (ALL) and the significance of detection of minimal residual disease (MRD), TCR Vδ 2 Dδ 3 rearrangement was detected in 40 cases of ALL by using nested polymerase chain reaction (PCR). The results showed that the clone gene rearrangement of TCR Vδ 2 Dδ 3 was found in 30 ALL (75%). The positive rate of TCR Vδ 2 Dδ 3 rearrangement was higher in the patients with B ALL (80%) than in those with T ALL ( P <0.01). MRD was converted into negative in 5 cases with continuous complete remission (CCR). Among the 5 cases, 4 terminated chemotherapy and survived for 85～199 months. Eight cases of MRD converted into weak positive survived for 41～187 months, and 3 of them experienced a bone marrow relapse and stayed in CR again. Now these 8 cases are all in continuous chemotherapy. In seven out of 13 cases of persisted MRD and of 1 case of reappearing MRD, the bone marrow relapsed and 5 of them died. The results suggested that the patients with CR over 3 years and with negative MRD had better prognosis. These could be considered as the index of terminating chemotherapy. When MRD persisted or reappeared, bone marrow relapse was possible and effective chemotherapy was needed. The results of this study showed that dynamic detection of MRD in ALL by using nested PCR played an important role in guiding clinical treatment, predicting bone marrow relapse and evaluating prognosis.