等位基因特异性聚合酶链反应结合测序对骨髓增殖性疾病JAK2 V617F点突变的研究

The investigation of JAK2 V617F point mutation in myeloproliferative disorders by allele-specific polymerase chain reaction in combination with sequence analysis

目的研究骨髓增殖性疾病(MPD)患者 Janus 激酶2(JAK2)V617F 点突变,探讨其临床应用意义。方法应用等位基因特异性聚合酶链反应(AS-PCR)结合测序检测20例慢性粒细胞白血病(CML)、23例真性红细胞增多症(PV)、40例原发性血小板增多症(ET)、8例慢性特发性骨髓纤维化(IMF)、3例高嗜酸粒细胞综合征(HES)患者外周血单个核细胞基因组 DNA 的 JAK2 V617F 点突变。结果 74例 BCR/ABL 阴性 MPD 患者中共发现38例存在 JAK2 V617F 突变(51.4%),分别为16例 PV,18例 ET,3例 IMF,1例 HES。对所有阳性样本和10例阴性样本进行测序鉴定,二者结果均一致。JAK2 V617F 突变阳性 ET 患者的外周血血红蛋白、红细胞压积和中性粒细胞比例显著高于阴性组。结论 JAK2 V617F 突变为 BCR/ABL 阴性 MPD 患者主要的分子遗传学异常,对其诊断、分类及探索有效的靶向治疗有重要影响。

Objective To study the JAK2 V617F point mutation in myeloproliferative disorders （MPD） and explore the clinical significance. Methods We used Allele-specific polymerase chain reaction （AS-PCR） in combination with sequence analysis to detect the mutation in genomic DNA of peripheral blood mononuclear cells from 20 chronic myelogenous leukemia （CML） patients, 23 polycythaemia vera （PV）, 40 essential thrombocythaemia （ET）, 8 idiopathic myelofibrosis （IMF）, 3 hypereosinophilic syndrome （HES）. Results JAK2 V617F was found in 38 （51.4%） of 74 BCR/ABL-negative MPD including 16 PV, 18 ET, 3 IMF and 1 HES patients. All positive samples and 10 negative samples identified by AS-PCR were confirmed by sequence analysis. Mutation-positive patients with ET had significantly increased hemoglobin, hematocrit, and neutrophil proportion than those without the mutation. Conclusion JAK2 V617F mutation is the key molecular genetics feature of BCR/ABL-negative MPD. Detection of JAK2 V617F mutation will bring about a major impact to the diagnosis, classification and treatment of MPD.