Two Novel Methods for Rapid Detection and Quantification of DNMT3A R882 Mutations in Acute Myeloid Leukemia
Two Novel Methods for Rapid Detection and Quantification of DNMT3A R882 Mutations in Acute Myeloid Leukemia
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DOI：
10.1016/j.jmoldx.2014.10.003
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发表时间：
2015-03-01
影响因子：
4.1
通讯作者：
Lo-Coco, Francesco
中科院分区：
文献类型：
--
作者：
Mancini, Melissa;Hasan, Syed Khizer;Lo-Coco, Francesco
DNMT 3A突变代表在具有正常核型的急性髓性白血病中可检测到的最常见的基因改变之一。虽然已经描述了DNMT 3A的各种复发性体细胞突变，但最常见的突变位于该基因的甲基转移酶结构域中的氨基酸R882处。据报道，DNMT 3A突变在疾病进展期间是稳定的，并且与具有正常核型的急性髓性白血病患者的不利结果相关。由于其预后意义和疾病演变过程中的高稳定性，DNMT 3A突变可能是微小残留疾病监测的高度信息化生物标志物。我们描述了一种新的快速诊断RT-PCR检测的基础上TauI限制性内切酶反应，以确定DNMT 3A R882突变的诊断。此外，我们还开发了一种基于肽核酸实时荧光PCR技术的灵敏度和特异性检测方法，用于监测DNMT 3A R882 H突变。我们从134例急性髓性白血病筛选样本中鉴定出24例DNMT 3A R882 H突变患者，并分析了这些患者在诱导和巩固治疗后微小残留病的动力学。该测定可用于更好地评估携带DNMT 3A R882 H突变的急性髓性白血病患者对治疗的反应。
DNMT3A mutations represent one of the most frequent gene alterations detectable in acute myeloid Leukemia with normal karyotype. Although various recurrent somatic mutations of DNMT3A have been described, the most common mutation is Located at amino acid R882 in the methyltransferase domain of the gene. DNMT3A mutations have been reported to be stable during disease progression and are associated with unfavorable outcome in acute myeloid Leukemia patients with normal karyotype. Because of their prognostic significance and high stability during disease evolution, DNMT3A mutations might represent highly informative biomarkers for minimal residual disease monitoring. We describe a new rapid diagnostic RT-PCR assay based on TauI restriction enzyme reaction to identify DNMT3A R882 mutations at diagnosis. In addition, we developed a sensitive and specific test based on peptide nucleic acid real-time PCR technology to monitor DNMT3A R882H mutation. We identified 24 DNMT3A R882H mutated patients out of 134 acute myeloid Leukemia screened samples and we analyzed in these patients the kinetics of minimal residual disease after induction and consolidation therapy. This assay may be useful to better assess response to therapy in patients with acute myeloid leukemia bearing the DNMT3A R882H mutation.