Drandi Daniela, Kubiczkova-Besse Lenka, Ferrero Simone, Dani Nadia, Passera Roberto, Mantoan Barbara, Gambella Manuela, Monitillo Luigia, Saraci Elona, Ghione Paola, Genuardi Elisa, Barbero Daniela, Omedè Paola, Barberio Davide, Hajek Roman, Vitolo Umberto, Palumbo Antonio, Cortelazzo Sergio, Boccadoro Mario, Inghirami Giorgio, Ladetto Marco
Division of Hematology, Department of Molecular Biotechnologies and Health Sciences, University of Torino, Torino, Italy.
Babak Myeloma Group, Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
J Mol Diagn. 2015 Nov;17(6):652-60. doi: 10.1016/j.jmoldx.2015.05.007. Epub 2015 Aug 28.
Real-time quantitative PCR (qPCR) is a well-established tool for minimal residual disease (MRD) detection in mature lymphoid malignancies. Despite remarkable sensitivity and specificity, qPCR has some limitations, particularly in the need for a reference standard curve, based on target serial dilutions. In this study, we established droplet digital PCR (ddPCR) for MRD monitoring in multiple myeloma, mantle cell lymphoma, and follicular lymphoma and compared it head-to-head with qPCR. We observed that ddPCR has sensitivity, accuracy, and reproducibility comparable with qPCR. We then compared the two approaches in 69 patients with a documented molecular marker at diagnosis (18 multiple myelomas, 21 mantle cell lymphomas assessed with the immunoglobulin gene rearrangement, and 30 follicular lymphomas with the use of the BCL2/immunoglobulin gene major breakpoint region rearrangement). ddPCR was successful in 100% of cases, whereas qPCR failed to provide a reliable standard curve in three patients. Overall, 222 of 225 samples were evaluable by both methods. The comparison highlighted a good concordance (r = 0.94, P < 0.0001) with 189 of 222 samples (85.1%; 95% CI, 80.4%-89.8%) being fully concordant. We found that ddPCR is a reliable tool for MRD detection with greater applicability and reduced labor intensiveness than qPCR. It will be necessary to authorize ddPCR as an outcome predictor tool in controlled clinical settings and multilaboratory standardization programs.
实时定量聚合酶链反应（qPCR）是一种成熟的用于检测成熟淋巴系恶性肿瘤微小残留病（MRD）的工具。尽管qPCR具有显著的灵敏度和特异性，但它也存在一些局限性，尤其是需要基于目标系列稀释建立参考标准曲线。在本研究中，我们建立了用于多发性骨髓瘤、套细胞淋巴瘤和滤泡性淋巴瘤MRD监测的液滴数字PCR（ddPCR），并将其与qPCR进行了直接比较。我们观察到ddPCR的灵敏度、准确性和可重复性与qPCR相当。然后，我们在69例诊断时有记录的分子标志物的患者中比较了这两种方法（18例多发性骨髓瘤、21例通过免疫球蛋白基因重排评估的套细胞淋巴瘤以及30例使用BCL2/免疫球蛋白基因主要断裂点区域重排的滤泡性淋巴瘤）。ddPCR在100%的病例中成功，而qPCR在3例患者中未能提供可靠的标准曲线。总体而言，225个样本中的222个可通过两种方法进行评估。比较结果显示两者具有良好的一致性（r = 0.94，P < 0.0001），222个样本中的189个（85.1%；95% CI，80.4% - 89.8%）完全一致。我们发现ddPCR是一种可靠的MRD检测工具，与qPCR相比具有更高的适用性和更低的劳动强度。有必要在受控的临床环境和多实验室标准化项目中授权将ddPCR作为一种结果预测工具。
