IG/TCR重排二代测序技术检测急性淋巴细胞白血病微小残留病的研究

Detection of minimal residual disease in acute lymphoblastic leukemia by next-generation sequencing of IG/TCR

目的:比较免疫球蛋白(IG)/T细胞受体(TCR)重排二代测序技术(next-generation sequencing,NGS)与其他急性淋巴细胞白血病(acute lymphoblastic leukemia,ALL)微小残留病(minimal residual disease,MRD)检测手段间的差异,评估IG/TCR重排NGS在ALL MRD监测中的价值。方法:回顾性分析2018年9月至2023年12月本院收治的25例ALL患者,同时采用NGS、流式细胞术(FCM)和实时定量聚合酶链反应技术(q-PCR)检测MRD。结果:48.0%的患者初诊时NGS检测存在3个或以上异常高频克隆。81.3%的患者治疗后的主要残留克隆与初诊时最高频克隆一致。在FCM判定MRD阴性的69例样本中24例(34.8%)经NGS检测仍阳性,MRD中位水平为4.13×10^(-5),显著低于FCM阳性患者的MRD水平(4.70×10^(-3))(P<0.05)。Ph+ALL患者同时采用NGS、q-PCR、FCM三种方法进行MRD检测,阳性率依次为59.1%、50.0%、27.3%,NGS阳性率显著高于FCM(P=0.035)。FCM判定阴性的16例样本中7例(43.8%)经NGS检测为阳性,中位MRD水平为1.99×10^(-5),较NGS/FCM双阳性组的中位MRD水平有降低趋势,但差异无统计学意义(P>0.05)。q-PCR判定阴性的11例样本中3例(27.3%)经NGS检测为阳性,中位MRD水平为7.59×10^(-6),较NGS/q-PCR双阳性组的中位MRD水平有降低趋势,但差异亦无统计学意义(P=0.053)。结论:NGS检测MRD的灵敏度和深度均优于FCM;Ph+ALL患者中NGS检测的灵敏度及深度较q-PCR有改善趋势。IG/TCR重排NGS是能保证足够检测深度的有效MRD检测手段。

Objective To compare the immunoglobulin(IG)/T cell receptor(TCR)rearrangement next-generation sequencing(NGS)with other methods of minimal residual disease(MRD)detection in acute lymphoblastic leukemia(ALL),and evaluating the value of IG/TCR rearrangement NGS in monitoring MRD.Methods A retrospective analysis was conducted on 25 ALL patients admitted to our hospital from September 2018 to December 2023.The MRD was determined by using NGS,flow cytometry(FCM)and real-time quantitative PCR(q-PCR).Results Three or more abnormal clones were detected by NGS in 48.0%of patients at initial diagnosis.The main residual clone of 81.3%of patients after treatment was consistent with the highest frequency clone at initial diagnosis.Among the 69 FCM MRD-negative(MRDneg)samples,24 cases(34.8%)were NGS MRD-positive(MRDpos),and the median MRD level was 4.13×10^(-5),which was significantly lower than that of FCM MRDpos patients(4.70×10^(-3))(P<0.05).In Ph+ALL patients,the positive rates of MRD detected by NGS,q-PCR and FCM were 59.1%,50.0%and 27.3%,respectively.The MRD positive rate of NGS was higher than that of FCM(P=0.035).Among the 16 FCM MRDneg samples,7 cases(43.8%)were NGS MRDpos.The median MRD level was 1.99×10^(-5),which had a decrease compared with the NGS/FCM MRDpos group,but there was no significant difference(P>0.05).Among the 11 q-PCR MRDneg samples,3 cases(27.3%)were NGS MRDpos,and the median MRD level was 7.59×10^(-6),which showed a decreasing trend compared with the NGS/q-PCR MRDpos group(P=0.053).Conclusion The MRD detection sensitivity and depth of NGS are better than that of FCM.NGS also showed advantages over q-PCR in the detection of sensitivity and depth in Ph+ALL patients.NGS of IG/TCR is an effective method to ensure sufficient MRD detection depth.