Er Tze-Kiong, Lin Sheng-Fung, Chang Jan-Gowth, Hsieh Li-Ling, Lin Shu-Kai, Wang Li-Hsuan, Lin Chin-Wen, Chang Chao-Sung, Liu Ta-Chih
Division of Molecular Diagnostics, Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan.
Clin Chim Acta. 2009 Oct;408(1-2):39-44. doi: 10.1016/j.cca.2009.07.002. Epub 2009 Jul 10.
Janus kinase 2 (JAK2) is a tyrosine kinase involved in the cytokine signaling of several growth factors such as erythropoietin and thrombopoietin in normal and neoplastic cells. The G to T exchange at nucleotide 1849 in exon 14 of the JAK2 gene leads to a substitution of valine with phenylalanine at the amino acid position 617 (V617F) of the JAK2 protein. Currently, the occurrence of the JAK2 V617F mutation is well recognized in chronic myeloproliferative disorders (MPDs).
We identified JAK2 V617F missense mutation in patients with MPD by high resolution melting (HRM) analysis. HRM analysis is a new gene scan tool that quickly performs the PCR and identifies sequences alterations without requiring post-PCR treatment. This study included 7 PV patients (41.1%), 6 ET patients (35.3%), and 4 myelofibrosis patients (23.5%). Additionally, our methodology was compared with amplification refractory mutation system (ARMS) assay.
Up to 5% of the JAK2 V617F mutation was successfully detected in patients with MPD using HRM analysis. Eleven out of 17 patients (64.7%) were positive for the presence of JAK2 V617F mutation. The prevalence of mutation in the different subtypes of MPDs was 85.7% in PV (6 of 7 patients), 66.7% in ET (4 of 6) and 5.9% in myelofibrosis (1 of 4). The results proved 100% comparable to those obtained by ARMS assay.
The HRM analysis is a rapid and effective technique for the detection of JAK2 V617F missense mutation.
Janus激酶2（JAK2）是一种酪氨酸激酶，参与正常细胞和肿瘤细胞中多种生长因子（如促红细胞生成素和血小板生成素）的细胞因子信号传导。JAK2基因第14外显子核苷酸1849处的G到T交换导致JAK2蛋白第617位氨基酸（V617F）处缬氨酸被苯丙氨酸取代。目前，JAK2 V617F突变在慢性骨髓增殖性疾病（MPD）中的发生已得到充分认识。
我们通过高分辨率熔解（HRM）分析在MPD患者中鉴定JAK2 V617F错义突变。HRM分析是一种新的基因扫描工具，可快速进行PCR并识别序列改变，无需PCR后处理。本研究纳入7例真性红细胞增多症（PV）患者（41.1%）、6例原发性血小板增多症（ET）患者（35.3%）和4例骨髓纤维化患者（23.5%）。此外，我们将该方法与扩增阻滞突变系统（ARMS）检测进行了比较。
使用HRM分析在MPD患者中成功检测到高达5%的JAK2 V617F突变。17例患者中有11例（64.7%）JAK2 V617F突变呈阳性。MPD不同亚型的突变发生率在PV中为85.7%（7例患者中的6例），在ET中为66.7%（6例中的4例），在骨髓纤维化中为5.9%（4例中的1例）。结果证明与ARMS检测获得的结果100%可比。
HRM分析是检测JAK2 V617F错义突变的一种快速有效的技术。
