Kotrova M, van der Velden V H J, van Dongen J J M, Formankova R, Sedlacek P, Brüggemann M, Zuna J, Stary J, Trka J, Fronkova E
CLIP Laboratory Centre, Department of Pediatric Hematology/Oncology, 2nd Faculty of Medicine and University Hospital Motol, Prague, Czech Republic.
Department of Immunology, Erasmus MC, University Medical Center Rotterdam, Rotterdam, The Netherlands.
Bone Marrow Transplant. 2017 Jul;52(7):962-968. doi: 10.1038/bmt.2017.16. Epub 2017 Feb 27.
Minimal residual disease (MRD) monitoring via quantitative PCR (qPCR) detection of Ag receptor gene rearrangements has been the most sensitive method for predicting prognosis and making post-transplant treatment decisions for patients with ALL. Despite the broad clinical usefulness and standardization of this method, we and others have repeatedly reported the possibility of false-positive MRD results caused by massive B-lymphocyte regeneration after stem cell transplantation (SCT). Next-generation sequencing (NGS) enables precise and sensitive detection of multiple Ag receptor rearrangements, thus providing a more specific readout compared to qPCR. We investigated two cohorts of children with ALL who underwent SCT (30 patients and 228 samples). The first cohort consisted of 17 patients who remained in long-term CR after SCT despite having low MRD positivity (<0.01%) at least once during post-SCT monitoring using qPCR. Only one of 27 qPCR-positive samples was confirmed to be positive by NGS. Conversely, 10 of 15 samples with low qPCR-detected MRD positivity from 13 patients who subsequently relapsed were also confirmed to be positive by NGS (P=0.002). These data show that NGS has a better specificity in post-SCT ALL management and indicate that treatment interventions aimed at reverting impending relapse should not be based on qPCR only.
通过定量聚合酶链反应（qPCR）检测抗原受体基因重排来监测微小残留病（MRD），一直是预测急性淋巴细胞白血病（ALL）患者预后和做出移植后治疗决策的最敏感方法。尽管该方法具有广泛的临床实用性且已标准化，但我们和其他研究团队多次报告称，干细胞移植（SCT）后大量B淋巴细胞再生可能导致MRD结果出现假阳性。新一代测序（NGS）能够精确且灵敏地检测多种抗原受体重排，因此与qPCR相比，能提供更具特异性的结果。我们调查了两组接受SCT的ALL患儿（30例患者，228份样本）。第一组包括17例患者，他们在SCT后长期处于完全缓解（CR）状态，尽管在SCT后监测期间至少有一次qPCR检测到的MRD阳性率较低（<0.01%）。在27份qPCR阳性样本中，只有1份经NGS确认为阳性。相反，在随后复发的13例患者中，15份qPCR检测到的MRD阳性率较低的样本中有10份也经NGS确认为阳性（P = 0.002）。这些数据表明，在SCT后的ALL管理中，NGS具有更好的特异性，并表明针对即将复发的治疗干预不应仅基于qPCR。
