JAK2V617F突变分型和相对定量研究——附123例慢性骨髓增殖性疾病检测结果分析  

Analysis for mutational status and relative quantitation of JAK2V617F——The detection result of JAK2V617F mutation in 123 patients with chronic myeloproliferative disorders.

作　　者：沈益民[1] 晁红颖[1] 张日[1] 冯宇峰[1] 岑建农[1] 姚利[1] 沈宏杰[1] 朱子玲[1] 薛永权[1] 

机构地区：[1]苏州大学附属第一人民医院血液科江苏省血液病研究所,江苏苏州215006

出　　处：《中国实用内科杂志》2009年第3期240-242,共3页Chinese Journal of Practical Internal Medicine

摘　　要：目的研究JAK2V617F突变在慢性骨髓增殖性疾病(CMPD)中的发生率和突变类型,并对突变转录本水平进行定量分析。方法对2003年1月至2008年5月苏州大学附属第一医院就诊的CMPD患者采用突变特异性扩增系统(ARMS)PCR方法检测JAK2V617F的发生率及其突变类型;采用毛细管电泳方法定量分析JAK2V617F突变转录本水平。结果123例CMPD患者共检出90例JAK2V617F阳性,其中35例真性红细胞增多症(PV)患者中JAK2V617F阳性率100%,85例原发性血小板增多症(ET)患者中JAK2V617F阳性率62.4%,低于PV患者,差异有统计学意义(P<0.05);3例慢性骨髓纤维化(IMF)患者中JAK2V617F阳性率66.7%。90例JAK2V617F突变患者中共检出纯合突变35例,其中PV患者17例(17/35),占48.6%,ET患者17例(17/85),占20.0%,低于PV患者,差异有统计学意义(P<0.05),IMF患者1例;毛细管电泳定量分析显示,纯合型突变患者JAK2V617F突变转录本水平较杂合型突变患者低,差异有统计学意义(P<0.05);杂合型PV患者JAK2V617F突变转录本水平高于杂和型ET患者,差异有统计学意义(P<0.05)。93例患者进行了染色体检查,6例有核型异常,但未发现特异性染色体改变。结论ARMS-PCR可作为JAK2V617F突变较准确的检测方法,结合毛细管电泳可用于此突变的定量分析以及临床CMPD的诊断。Objective To investigate the frequency and mutational status of JAK2V617F mutation in patients with chronic myeloproliferative disorders（CMPD） and to study the relative quantitation of mutated JAK2 mRNA. Methods Between January 2003 and May 2008,123 patients with MPD, who visited the First Affiliated Hospital of Soochow University, were enrolled in our study. JAK2V617F mutation and the mutational status were screened with amplification-refractory mutation sequencing polymerase chain reaction（ARMS-PCR） ;the relative quantation of mutated JAK2 mRNA was studied by using capillary electrophoresis. Results JAK2V617F mutation was detected in 35 （ 100% ） of the 35 patients with PV, 53 （62. 4% ）of the 85 with ET and 2（66. 7% ） of the 3 with IMF,tbe difference between PV and ET was significant（P 〈 0. 05）. Of 90 JAK2V617F patients examined,17（48.6% ,17/35） patients with PV and 17 （20% ,17/85） patients with ET and 1 （50%, 1/2 ） patient with IMF were homozygotes. ET patients showed lower prevalence of homozygote （P 〈 0. 05）. A quantitative assay by capillary electrophoresis showed that the mutated mRNA ratio was higher in the JAK2V617F homozygote in the JAK2V617F heterozygote patients. 18 PV heterozygote patients showed higher levels of mutated JAK2 mRNA than 36 heterozygote ET patients（ P 〈 0. 05 ）. Cytogenetic analysis was performed in 93 of the 123 patients,6 patients exhibited abnormal karyotype, but special chromosomal abnormality were not found. Conclusion The ARMS PCR technique can be used to detect the frequency and mutational status of JAK2V617F mutation owing to sensitivity. Along with capillary electrophoresis,a quantitative assay for mutated JAK2 mRNA,the diagnosis of CMPD become easy.

关 键 词：慢性骨髓增殖性疾病 JAK2V617F ARMS—PCR 毛细管电泳 

分 类 号：R5[医药卫生—内科学]