JAK2 V617F 突变可通过液滴数字 PCR 可靠地检测到血清中。
JAK2 V617F mutation can be reliably detected in serum using droplet digital PCR.
机构信息
Centre for Laboratory medicine, Østfold Hospital Trust, Kalnes, Norway.
Department of Oncology, Østfold Hospital Trust, Kalnes, Norway.
出版信息
Int J Lab Hematol. 2018 Apr;40(2):181-186. doi: 10.1111/ijlh.12762. Epub 2017 Nov 17.
INTRODUCTION
Detection of the JAK2 V617F mutation is a key step in the diagnosis of myeloproliferative neoplasms (MPN). Sensitive real-time quantitative PCR (qPCR) detection on peripheral blood (PB) is the most widely used method. The main objective of this study was to determine whether serum, the most common material available in archival biobanks, is a good liquid biopsy for detecting and quantifying the JAK2 V617F mutation using droplet digital PCR (ddPCR).
METHODS
Paired PB and serum samples from 66 patients with MPN were used. Serum samples were frozen at -25°C before analysis. DNA was extracted from 200 μL PB and 400 μL serum, and ddPCR analysis was performed.
RESULTS
Among the 47 patients with detectable mutation in their PB samples, the overall sensitivity for the detection of JAK2 mutation in serum was of 96% (45 of 47); V617F was detected in all cases where mutation load was above 1%. Our results showed very strong correlation between PB and serum (Spearman r: 0.989, P < .0001). Significantly higher allele burden was detected in serum compared to PB (Wilcoxon signed ranks test, Z = -5.672, P < .0001).
CONCLUSION
In our study, JAK2 V617F mutation load as low as 1% was reliably detected in serum using ddPCR.
简介
检测 JAK2 V617F 突变是骨髓增殖性肿瘤（MPN）诊断的关键步骤。外周血（PB）的敏感实时定量 PCR（qPCR）检测是最常用的方法。本研究的主要目的是确定血清（存档生物库中最常见的可用材料）是否可作为检测和定量 JAK2 V617F 突变的良好液体活检方法，方法是使用液滴数字 PCR（ddPCR）。
方法
使用 66 例 MPN 患者的配对 PB 和血清样本。在分析前将血清样本在-25°C 下冷冻。从 200μL PB 和 400μL 血清中提取 DNA，并进行 ddPCR 分析。
结果
在 47 例 PB 样本中可检测到突变的患者中，血清中 JAK2 突变检测的总灵敏度为 96%（45/47）；在突变载量高于 1%的所有情况下均检测到 V617F。我们的结果显示 PB 和血清之间具有非常强的相关性（Spearman r：0.989，P<.0001）。与 PB 相比，血清中检测到的等位基因负担明显更高（Wilcoxon 符号秩检验，Z=-5.672，P<.0001）。
结论
在我们的研究中，使用 ddPCR 可可靠地在血清中检测到低至 1%的 JAK2 V617F 突变载量。
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