文章：

传统PCR与下一代测序在急性髓系白血病FLT3、IDH及NPM1突变诊断中的比较：PETHEMA PCR-LMA研究结果

Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study

原文发布日期：1 March 2025

DOI: 10.3390/cancers17050854

类型: Article

开放获取: 是

英文摘要：

Background/Objectives: This PETHEMA PCR-LMA study aimed to evaluate whether mutations detected by NGS (VAF cut-off of ≥5%) correlate with NPM1, FLT3-ITD, FLT3-TKD, IDH1, and IDH2 mutations detected using conventional PCR (analytical sensitivity 3%) in a nationwide network of seven reference laboratories. Methods: Between 2019 and 2021, 1685 adult AML patients with at least one centralized sample (NGS or PCR) at primary diagnosis or relapse/refractory episode were included. Results: During this period, 1288 paired NGS/PCR samples (1094 at diagnosis, 103 at relapse and 88 at refractoriness) were analyzed. Considering PCR the gold-standard, for NPM1 NGS sensitivity was 98.5% and specificity 98.9%, for FLT3-ITD 73.8% and 99.6%, for FLT3-TKD 84.5% and 99.3%, for IDH1 98.7% and 98.7%, and for IDH2 99.1% and 97.7%, respectively. Overall concordance rate of positive results between NGS (and PCR was 95% (262/276) for NPM1, 72% (149/206) for FLT3-ITD, 74% (49/66) for FLT3-TKD, 87% (77/89) for IDH1 and 84% (107/127) for IDH2. Overall, median days from sample reception until report were 7 for PCR and 28 for NGS. Conclusions: This study shows high concordance between NPM1 and IDH results using PCR and NGS. However, sensible important discrepancies are observed for FLT3 mutations. In our context, rapid screening for these druggable mutations should be performed by conventional PCR.

摘要翻译： 

背景/目的：本研究旨在通过PETHEMA PCR-LMA项目，评估在由七家参考实验室组成的全国性检测网络中，采用二代测序技术（突变等位基因频率阈值设定为≥5%）检测出的突变，与传统PCR方法（分析灵敏度为3%）检测NPM1、FLT3-ITD、FLT3-TKD、IDH1及IDH2突变的结果之间是否存在相关性。方法：研究纳入了2019年至2021年间1685例成年急性髓系白血病患者，这些患者在初诊或复发/难治阶段至少提供了一份集中检测样本（用于二代测序或PCR检测）。结果：在此期间，共分析了1288对匹配的二代测序/PCR样本（其中1094份为初诊时样本，103份为复发时样本，88份为难治时样本）。以PCR检测结果为金标准，二代测序检测NPM1突变的敏感性和特异性分别为98.5%和98.9%，FLT3-ITD为73.8%和99.6%，FLT3-TKD为84.5%和99.3%，IDH1为98.7%和98.7%，IDH2为99.1%和97.7%。二代测序与PCR检测阳性结果的总体一致率分别为：NPM1 95%（262/276），FLT3-ITD 72%（149/206），FLT3-TKD 74%（49/66），IDH1 87%（77/89），IDH2 84%（107/127）。总体而言，从样本接收到报告发出的中位时间，PCR检测为7天，二代测序为28天。结论：本研究表明，采用PCR和二代测序检测NPM1和IDH突变的结果具有高度一致性。然而，在FLT3突变检测中观察到显著差异。在本研究背景下，针对这些可靶向治疗的突变进行快速筛查，仍应通过传统PCR方法完成。

原文链接：

Conventional PCR Versus Next Generation Sequencing for Diagnosis of FLT3, IDH and NPM1 Mutations in Acute Myeloid Leukemia: Results of the PETHEMA PCR-LMA Study