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目的利用六色流式细胞术(FCM)检测急性B淋巴细胞白血病(B-ALL)微小残留病变(MRD),探讨传统用于临床危险度分级各因素与六色FCM检测MRD值间关系及化疗不同时间点MRD检测值间关系。方法研究对象为2010-7-15—2011-1-25在广州市妇女儿童医疗中心初诊并接受正规化疗的25例B-ALL患儿。根据患儿初发时免疫分型结果,分别在诱导化疗第15、33天及大剂量MTX化疗前3个时间点进行MRD检测,每例患儿使用一组六色同型对照及根据初诊时免疫分型结果灵活调整的两组六色抗体组合进行检测。一组六色同型对照为IgG1-FITC/IgG1-PE/IgG1-PE-Cy7/IgG1-APC/IgG1-Percp/IgG1-APC-cy7;两组六色抗体组合为CD20-FITC/CD34-PE/CD10-PE-Cy7/CD22-APC/CD45-Percp/CD19-APC-cy7、CD7-FITC/CD5-PE/CD13-PE-Cy7/CD33-APC/CD45-Percp/CD19-APC-cy7或CD38-FITC/CD34-PE/CD10-PE-Cy7/CD22-APC/CD45-Percp/CD19-APC-cy7、CD7-FITC/CD15-PE/CD34-PE-Cy7/CD33-APC/CD45-Percp/CD19-APC-cy7。分析各时间点MRD检测值与传统用于临床危险度分级各因素间关系及3个不同检测时间点MRD检测值间关系。结果化疗第15天使用六色FCM检出骨髓MRD阳性20例,P25为0.19%,P50为0.67%,P75为4.21%;第33天检出骨髓MRD阳性10例,P25为0.07%,P50为0.10%,P75为0.26%;大剂量MTX化疗前检出骨髓MRD阳性14例,P25为0.03%,P50为0.04%,P75为0.09%。结论在诱导化疗阶段、诱导化疗结束时MRD的阳性率与诱导化疗第15天相比有显著下降;大剂量MTX化疗前MRD阳性值与诱导化疗第15天和诱导化疗结束时相比,有随化疗时间延长逐步下降的趋势。按传统危险度分级的相关因素分别分组,各组在诱导化疗第15天、诱导化疗结束时和大剂量MTX化疗前MRD检出率均无明显差异。
Objective To detect the minimal residual disease (MRD) of acute lymphoblastic leukemia (B-ALL) by six-color flow cytometry (FCM), and to explore the relationship between the MRD of traditionally used in clinical risk classification and six-color FCM The relationship between MRD values at different time points. Methods The subjects were 25 children with B-ALL who were newly diagnosed at Guangzhou Women and Children’s Medical Center and who underwent formal chemotherapy during 2010-7-15-2011-1-25. According to the results of immunophenotyping in the first episode, MRD was performed on the 15th and the 33th day of induction chemotherapy and 3 times before the high dose MTX chemotherapy. One case of six-color isotype control and the first immunization The typing results were flexibly adjusted for the detection of two groups of six-color antibodies. A group of six-color isotype control was IgG1-FITC / IgG1-PE / IgG1-PE-Cy7 / IgG1-APC / IgG1-Percp / IgG1-APC- CD7-FITC / CD5-PE / CD13-PE-Cy7 / CD33-APC / CD45-Percp / CD19-APC-cy7 or CD38- FITC / CD34-PE / CD10-PE-Cy7 / CD22-APC / CD45- Percp / CD19- APC- cy7, CD7- FITC / CD15- PE / CD34- PE- Cy7 / CD33- APC / CD45- Percp / APC-cy7. The relationship between the MRD values at various time points and the traditional factors used for clinical risk grade and the MRD values at three different time points were analyzed. Results On the 15th day of chemotherapy, 20 cases with positive MRD were detected by six color FCM. P25 was 0.19%, P50 was 0.67% and P75 was 4.21%. On the 33rd day, bone marrow MRD was positive in 10 cases, P25 was 0.07%, P50 was 0.10%, and P75 was 0.26%. In the high-dose MTX group, 14 cases with positive MRD were detected in the bone marrow, P25 was 0.03%, P50 was 0.04% and P75 was 0.09%. Conclusions The positive rate of MRD at the end of induction chemotherapy is significantly lower than that of induction chemotherapy at the end of induction chemotherapy. Compared with the 15th day of induction chemotherapy and the end of induction chemotherapy, the positive rate of MRD before high dose MTX chemotherapy Chemotherapy time extended gradually declining trend. According to the traditional risk classification, the incidence of MRD was not significantly different between the groups on the 15th day of induction chemotherapy, the end of induction chemotherapy and the high dose MTX chemotherapy.