JAK2V617F基因突变的定量测定及在慢性骨髓增殖性疾病中的应用

Quantitative determination of JAK2V617F mutation and its application in chronic myeloproliferative disorders

目的探讨荧光定量PCR的方法检测JAK2V617F基因突变并分析其在慢性骨髓增殖性疾病(CMPD)中的诊断价值。方法采用荧光定量聚合酶链反应(PCR)检测68例确诊CMPD患者骨髓标本中JAK2V617F基因突变的阳性率和相对定量,分析其在CMPD中的诊断价值以及和临床资料之间的联系。结果 68例确诊患者的骨髓标本均成功扩增JAK2V617F野生型和突变型。JAK2V617F基因突变的总阳性率为66.2%(45/68),其中真性红细胞增多症(PV)患者阳性率75.0%(21/28),原发性血小板增多症(ET)阳性率60.6%(20/33),特发性骨髓纤维化(IMF)阳性率57.1%(4/7)。PV患者JAK2V617F突变阳性率明显高于ET和IMF患者。PV患者JAK2V617F的相对定量和血红蛋白数和白细胞数呈正相关。ET患者中JAK2V617F的相对定量和患者的血小板计数呈正相关。结论荧光定量PCR的方法能够快速准确的检测JAK2V617F基因突变及其突变比例,为临床诊断CMPD提供有效的实验室指标,JAK2V617F的相对定量和患者的临床血液学指标存在一定关系。

Objective To quantitatively determine JAK2V617F mutation by fluorescence quantitative polymerase chain reaction （FQ-PCR）and to analyze its diagnostic value in chronic myeloproliferative disorders（CMPD）.Methods FQ-PCR was adopted to detect the positive rate and the relative quantification of JAK2V617F gene mutation in the marrow samples of 68 patients with CMPD.Their diagnostic value in CMPD and their relation with the clinical data were analyzed.Results The marrow samples in 68 cases of CMPD were successfully amplified in wild-type and mutant JAK2V617F.The total positive rate of the JAK2V617F gene mutation was 66.2%（45/68）.The positive rate was 75.0%（21/28）in the patients with polycythemia vera（PV）,60.6%（20/33）in the patients with primary thrombocythemia（ET）and 57.1%（4/7）in patients with idiopathic myelofibrosis respectively.The posi-tive rate of JAK2V617F mutation in the PV patients was significantly higher than that in the patients with ET and IMF.The JAK2V617F relative quantification in the PV patients was positively correlated with the hemoglobin concentration and white blood cell（WBC）count,and which in the ET patients was positively correlated with the platelet count.Conclusion FQ-PCR can quickly and accurately detect JAK2V617F mutation and its mutation ratio,which provides an effective laboratory indicator for the clinical diagnosis of CMPD.Certain relation exists between the JAK2V617F relative quantification and the clinical hematological indexes of the patients.