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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 incidence and types of mutations in JAK2V617F mutation in chronic myeloproliferative disorders (CMPD) and to quantitatively analyze the level of the mutant transcripts. Methods The incidence and mutation types of JAK2V617F in patients with CMPD treated in the First Affiliated Hospital of Soochow University from January 2003 to May 2008 were detected by ARMS PCR. Capillary electrophoresis was used to quantitatively analyze the expression of JAK2V617F Mutant transcript levels. Results A total of 90 JAK2V617F positive cases were detected in 123 patients with CMPD. The positive rate of JAK2V617F was 100% in 35 patients with polycythemia vera (PV) and 62.4% in 85 patients with essential thrombocythemia (ET) The difference was statistically significant (P <0.05). The positive rate of JAK2V617F in 3 patients with chronic myelofibrosis (IMF) was 66.7%. Totally 35 homozygous mutations were detected in 90 patients with JAK2V617F mutation, of which 17 patients (17/35) were PV patients (48.6%), 17 patients (17/85) were ET patients, accounting for 20.0%, which was lower than PV patients Statistical analysis (P <0.05) showed that there was 1 IMF patient. Capillary electrophoresis quantitative analysis showed that the JAK2V617F mutation transcript level in patients with homozygous mutation was lower than that in heterozygous mutation group, with significant difference (P <0.05); The level of JAK2V617F mutation transcripts in syncytial PV patients was significantly higher than that in heterozygous ET patients (P <0.05). Ninety-three patients had a chromosomal test, and six had karyotype abnormalities, but no specific chromosomal alterations were found. Conclusion ARMS-PCR can be used as a more accurate method for detecting JAK2V617F mutation. Combined with capillary electrophoresis, it can be used for the quantitative analysis of this mutation and the diagnosis of clinical CMPD.