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探索应用TCRr基因重排标志及改良的极限稀释定量PCR法检测小儿ALL微小残留病(MRD)的可行性,探讨其追踪骨髓MRD变化的临床意义。方法:改良极限稀释定量PLR法。结果:敏感度平均为4拷贝。对14例初治阳性病人在诱导达到缓解(CR)时的MRD定量检测结果显示,骨髓复发组MRD定量值平均为0.343%,未复发组为0.004%(P<0.01)。14例中6例CR时MRD阴性,4例在追踪期内持续阴性,2例转为阳性(分别为0.017%、0.020%),化疗后MRD渐减少至转阴;3例CR时MRD平均值为0.077%,在缓解期MRD值渐减少,其中2例临床持续缓解,1例骨髓无复发但发生睾丸白血病,1例CR时MRD值较高,2例持续高值,2例MRD值渐增高,此5例均复发。结论:CR时MRD阳性的病人应持续监测其水平,若持续高值或逐渐增高,复发的可能性大,应加强化疗以防复发。采用极限稀释定量法检测ALL的MRD有简单、敏感性高的优点。
To explore the feasibility of using TCRr gene rearrangement markers and modified limit-dilution quantitative PCR to detect minimal residual disease (MRD) in children with ALL, and to explore its clinical significance in tracking changes of MRD in bone marrow. Methods: Modified limit dilution quantitative PLR method. Results: The average sensitivity was 4 copies. The quantitative MRD results of 14 newly diagnosed positive patients after induction of remission (CR) showed that the MRD quantitative value of bone marrow recurrence group was 0.343% on average and 0.004% on non-recurrence group (P <0.01) . Of the 14 cases, MRD was negative in 6 cases, persistent negative in 4 cases, positive in 2 cases (0.017%, 0.020% respectively), and MRD decreased gradually to negative after chemotherapy. Three cases of CR The average MRD was 0.077%. MRD decreased gradually in the remission period, of which 2 cases sustained clinical remission, 1 case had no recurrence of bone marrow but had testicular leukemia, 1 case had higher MRD and 2 sustained high values, 2 cases of MRD increased gradually, all 5 cases of recurrence. CONCLUSIONS: Patients with MRD positive at CR should be continuously monitored for their CR level. If they persist high or gradually increase, they are more likely to relapse and chemotherapy should be intensified to prevent relapse. The use of limit dilution quantitative detection of ALL MRD has the advantages of simple, high sensitivity.