JAK2V617F突变在诊断真性红细胞增多症的应用研究  

The clinic study of JAK2V617F mutation detection in diagnosis of polycythemia vera

作　　者：胡国瑜[1] 张玲[2] 袁朝晖[2] 罗丹[2] 张广森[1] 

机构地区：[1]中南大学湘雅二医院血液科,湖南长沙410011 [2]湖南省株洲市一医院血液科

出　　处：《中国医师杂志》2008年第9期1185-1188,共4页Journal of Chinese Physician

摘　　要：目的探索JAK2V617F突变在真性红细胞增多症诊断与鉴别诊断中意义。方法对38例确诊为真性红细胞增多症患者、20例继发性红细胞增多症的患者和20例正常健康对照者的外周血中性粒细胞基因组DNA进行提取，并应用等位基因PCR技术和PCR产物直接序列测定对-JAK2基因第14号外显子第1849位核苷酸是否存在G—T突变，以确定JAK2V617F突变诊断真性红细胞增多症的特异性，同时比较二种检测方法的敏感性和特异性。结果应用PCR产物序列测定的方法在38例真性红细胞增多症有32例（占84％），在20例继发性红细胞增多症和20例正常健康对照组中均发现一例突变，而用等位基因特异性PCR技术在38例真性红细胞增多症检测到35例（占92％）存在JAK2V617F突变，并经过PCR产物正反序列测定得到证实，在20例继发性红细胞增多症和20例正常健康对照组中均发现一例突变。结论JAK2V617F突变的检测可作为诊断为真性红细胞增多症的特异性分子标志物，而等位基因特异性PCR技术在检测JAK2V617F突变中较PCR产物序列测定具有更高的敏感性和特异性，更适合于临床实验室常规开展。Objective To investigate the diagnostic value of JAK2V617F mutation detection in the diagnosis of polycythemia vera, and screen a more simple method for clinic lahoratory to detect the mutation of JAK2V617F. Method DNA was extracted by standard procedures after isolating total leukocytes from peripheral blood mononuclear cells by density gradient centrifugation over Histopaque 1077. DNA samples were amplified, and single-stranded biotinylated PCR products were prepared for sequencing. At the same time, in order to testify the reliability of the allele-specific PCR, two forward primer and one common reverse primers were applied for identifying the mutation. Result In 32 of 38 patients with polycythemia vera, JAK2V617F mutation was determined by conventional DNA sequencing. 35 JAK2V617F mutations were detected by the allele-specific PCR, and these mutations were confirmed by DNA sequencing. None mutation was found in secondary polycythemia and normal control. Conclusion JAK2V617F function mutation occurs in nearly all patients with PV, and JAK2V617F mutation should be a molecular marker in the diagnosis of polycythemia vera. Allele-specific PCR is a very sensitive and specific method for detecting JAK2V617F mutation.

关 键 词：蛋白质酪氨酸激酶 点突变 红细胞增多症 真性/诊断 

分 类 号：R555.1[医药卫生—血液循环系统疾病] R699.2[医药卫生—内科学]