Department of Hematology Diagnostics, The University Hospital, Kraków, Poland.
Department of Pathology, Zealand University Hospital, Roskilde, Denmark.
Ann Hematol. 2018 Dec;97(12):2299-2308. doi: 10.1007/s00277-018-3451-1. Epub 2018 Jul 28.
Philadelphia-negative myeloproliferative neoplasms (MPNs) are a diverse group of diseases whose common feature is the presence of V617F mutation of the JAK2 gene. In the era of novel therapeutic strategies in MPNs, such as JAK-inhibitor therapy, there is a growing need for establishing high sensitive quantitative methods, which can be useful not only at diagnosis but also for monitoring therapeutic outcomes, such as minimal residual disease (MRD). In this study, we compared the qPCR and ddPCR methods and their clinical utility for diagnosis, prognostication, and treatment monitoring of MPNs with JAK2 V617F mutation in 63 MPN patients of which 6 were subjected to ruxolitinib treatment. We show a high conformance between the two methods (correlation coefficient r = 0.998 (p < 0.0001)). Our experiments revealed high analytical sensitivity for both tests, suggesting that they are capable of detecting the JAK2 V617F mutation at diagnosis of MPN with a limit of detection (LoD) of 0.12% for qPCR and 0.01% for ddPCR. The alterations of JAK2 V617F allele burden in patients treated with ruxolitinib were measured by both methods with equal accuracy. The results suggest an advantage of ddPCR in monitoring MRD because of allele burdens below the LoD of qPCR. Overall, the clinical utility of qPCR and ddPCR is very high, and both methods could be recommended for the routine detection of the V617F mutation at diagnosis, though ddPCR will probably supersede qPCR in the future due to cost-effectiveness.
费城阴性骨髓增殖性肿瘤（MPN）是一组具有不同特征的疾病，其共同特征是存在 JAK2 基因 V617F 突变。在 MPN 的新型治疗策略时代，如 JAK 抑制剂治疗，越来越需要建立高敏感定量方法，这些方法不仅在诊断时有用，而且对于监测治疗结果（如微小残留疾病（MRD））也很有用。在这项研究中，我们比较了 qPCR 和 ddPCR 方法及其在 63 名 JAK2 V617F 突变 MPN 患者中的临床应用，其中 6 名患者接受了鲁索替尼治疗。我们表明两种方法之间具有高度一致性（相关系数 r=0.998（p<0.0001））。我们的实验表明两种测试均具有很高的分析灵敏度，表明它们能够在 MPN 的诊断中检测到 JAK2 V617F 突变，qPCR 的检测限（LoD）为 0.12%，ddPCR 的检测限为 0.01%。用两种方法均可准确测量接受鲁索替尼治疗的患者的 JAK2 V617F 等位基因负担的改变。结果表明，ddPCR 在监测 MRD 方面具有优势，因为 qPCR 的 LoD 以下的等位基因负担。总体而言，qPCR 和 ddPCR 的临床应用非常高，并且两种方法都可以推荐用于常规检测诊断时的 V617F 突变，尽管由于成本效益，ddPCR 可能会在未来取代 qPCR。
