急性淋巴细胞白血病微小残留病不同监测方法比较  

Comparison of different monitoring methods of minimal residual disease in acute lymphoblastic leukemia

作　　者：徐岳一[1] 杨永公[1] 欧阳建[1] Xu Yueyi;Yang Yonggong;Ouyang Jian(Department of Hematology,the Affiliated Drum Tower Hospital of Nanjing University Medical School,Nanjing 210008,China)

机构地区：[1]南京大学医学院附属南京鼓楼医院血液科,210008

出　　处：《白血病．淋巴瘤》2020年第10期577-580,共4页Journal of Leukemia & Lymphoma

摘　　要：急性淋巴细胞白血病(ALL)患者的微小残留病(MRD)水平与预后密切相关。目前国内外通用的三种ALL-MRD检测手段包括免疫球蛋白重链/T细胞受体(IGH/TCR)基因重排检测法、流式细胞术(FCM)检测法、白血病相关融合基因检测法。IGH/TCR基因重排检测法包括实时荧光定量聚合酶链反应(RT-qPCR)和二代测序(NGS),前者主要检测IGH/TCR重排基因的可变区,比普通FCM灵敏一个对数级,但初诊时某些微小克隆易被忽略,导致假阴性;后者同样是检测IGH/TCR重排基因的可变区,其灵敏度较FCM及RT-qPCR法更高,最高可达10-6,并可以追踪导致复发的小亚克隆。FCM检测MRD的灵敏度通常在10-4,8色以上FCM可达10-6,但要求细胞数达(2~5)×107,而缓解期标本细胞数一般很难达到要求,且对检测人员技术要求较高,易受克隆演变和表型漂移的干扰。RT-qPCR可用于检测BCR-ABL等融合基因,灵敏度最高可达约10-5,但仅有少数患者存在可用作MRD监测的特异性融合基因改变。对于费城染色体阳性ALL患者推荐RT-qPCR法检测MRD水平,而对于费城染色体阴性ALL和T细胞ALL患者,FCM、RT-qPCR、NGS均适用。The level of minimal residual disease(MRD)is closely associated with prognosis in patients with acute lymphoblastic leukemia(ALL).Currently,3 kinds of ALL-MRD detection methods commonly used at home and abroad include immunoglobulin heavy chain and T-cell receptor(IGH/TCR)gene rearrangement assessment,flow cytometry(FCM)and leukemia-associated fusion gene detection.IGH/TCR gene rearrangement assessment methods include real-time fluorescent quantitative polymerase chain reaction(RT-qPCR)and next-generation sequencing(NGS).RT-qPCR mainly detects the variable region of IGH/TCR rearrangement genes;it is about one log more sensitive than FCM,but microclones may be easily ignored leading to false negative results.NGS also detects the variable region of IGH/TCR rearrangement genes.The sensitivity of NGS-based MRD assays is higher than that of FCM and RT-qPCR,and its sensitivity is up to 10-6,while small subclones causing recurrence can be tracked.The sensitivity of MRD was 10-4 detected by using FCM,while FCM with≥8-color can achieve 10-6.However,such high level of sensitivity requires(2-5)×107 nucleated cells,which is rarely obtainable from remission marrows.FCM also requires substantial expertise on inspectors,and results may be easily affected by clonal evolution or phenotype shift.RT-qPCR can be used to detect fusion genes such as BCR-ABL,with a sensitivity of up to about 10-5,but only few ALL patients carry specific gene fusions change that can be used as the monitoring of MRD.For Philadelphia chromosome-positive ALL patients,RT-qPCR is recommended to detect the level of MRD.For Philadelphia chromosome-negative ALL and T-cell ALL patients,FCM,RT-qPCR and NGS methods are all applicable.

关 键 词：白血病 淋巴细胞 急性 微小残留病 二代测序 

分 类 号：R733.71[医药卫生—肿瘤]