【作者】 艾国；

【导师】 刘昕； 陈新年；

【作者基本信息】 兰州大学 ， 病理学与病理生理学， 2009， 硕士

【摘要】 目的:对青海省bcr/abl融合基因阴性的骨髓增殖性疾病(myeloproliferativedisorders,MPD)病人进行JAK2V617F点突变定性检测,分析JAK2V617F点突变在我国西部地区MPD病人中的表达率,为临床诊断、治疗提供理论依据。方法:1.常规提取全基因组DNA、骨髓组织总RNA提取。2.用逆转录聚合酶链反应(RT-PCR)对140例MPD病人进行bcr/abl融合基因检测。3.用等位基因特异性聚合酶链反应(AS-PCR)检测140例MPD病人JAK2基因的V617F点突变,对有突变的病例进行测序验证。4.分析比较青海省与国内部分其它地区MPD病人JAK2基因的V617F点突变表达率的差异。结果:1、140例典型MPD病人bcr/abl融合基因检测:76例CML病人均为bcr/abl阳性的病人,其余64例典型MPD病人为bcr/abl阴性,其中32例为真性红细胞增多症,20例为原发性血小板增多症,12例为特发性骨髓纤维化病人。2、AS-PCR检测JAK2V617F突变发生率:bcr/abl阳性的76例慢性粒细胞白血病病人JAK2V617F突变均为阴性;64例bcr/abl阴性MPD病人中的突变率是:32例真性红细胞增多症病人中24例阳性(占75%),20例原发性血小板增多症病人中6例阳性(占30%),12例特发性骨髓纤维化病人中6例阳性(占50%)。3、青海省与国内部分其它地区MPD病人JAK2基因的V617F点突变率相比存在差别。结论:JAK2V617F点突变在青海省bcr/abl融合基因阴性的骨髓增殖性疾病的发病中起了重要作用,PV、ET、IMF病人JAK2V617F突变阳性率依次为75%(24/32)、30%(6/20)和50%(6/12),与国内其它部分地区相比存在差别。JAK2V617F可能为PV、ET、IMF的特征性分子事件,结合其临床病理特征可作为一个独立的分子指标用于此三类疾病的临床诊断。更多还原

【Abstract】 Objective:To detect the JAK2V617F mutation in myeloproliferative disorders(MPD) of Qinghai province by allele-specific primer polymerase chain reaction and evaluate the significance of JAK2V617F in diagnosis and therapy of MPD.Methods:1.Genomic DNA and total RNA were extracted from white blood cells and marrow respectively by route way.2.The bcr/abl fusion gene was detected in 140 Patients with MPD by the reverse transcription Polymerase chain reaction(PT-PCR).3.The JAK2V617F mutation was detected in 70 patients with MPD by allele-specific polymerase chain reaction(AS-PCR) and the positive cases with the mutation were checked by sequencing.Results:Between 140 Patients with MPD,the bcr/abl fusion gene is detected in 76 chronic myelocytic leukemia patients,others is negative,including 32 polycythemia vera,20 essential thrombocythemia,12 idiopathic myelofibrosis.The JAK2V617F mutation was not detected in 76 chronic myelocytic leukemia patients,whose bcr/abl fusion is negative.But in 24 of 32(75%) patients with polycythemia vera,6 of 20(30%) with essential thrombocythemia,6 of 12(50%) with idiopathic myelofibrosis.Conclusion:The difference of mutation frequencies among polycythemia vera,essential thrombocythemia and idiopathic myelofibrosis patients was of no statistical significance(P＞0.05).The JAK2V617F may be a characteristic molecular event in polycythemia vera,essential thrombocythemia and idiopathic myelofibrosis patients.It may serve as an important molecμlar marker for the diagnosis and classification of these three diseases.更多还原