CAJ | 학술논문

目的：建立多重等位基因特异性聚合酶链反应检测 JAK 2基因5种常见突变的方法，并评价在3种常见的骨髓增殖性疾病中的临床意义。方法对149例 BCR-ABL 融合基因为阴性的骨髓增殖性疾病（MPD）患者，包括73例真性红细胞增多症（PV）、51例原发性血小板增多症（ET）和25例原发性骨髓纤维化（IMF）患者；并结合临床及实验室其他检查资料，对其在该类疾病诊断和鉴别诊断中的价值进行评估。20名急性白血病患者和15名健康志愿者作为对照组进行研究。所有病例的骨髓或外周血标本抽提 DNA 后用建立的多重 PCR 方法进行平行检测。结果①149例 MPD 患者中共检出118例患者存在 JAK2基因突变，总突变率为79．2％。其中 PV 患者阳性68例，JAK2 V617F 突变65例、JAK2 exon12突变3例，阳性率93．2％（68/73）；ET 患者检测出阳性35例，其中34例为 V617F 突变，JAK2 exon12突变1例，阳性率为68．6（35/51）；IMF 患者阳性15例，均为 JAK2 V617F 突变，阳性率为60％（15/25）。②JAK2突变阳性的 MPD 患者和突变阴性 MPD 的临床资料相比较，两组在患者性别，发病年份方面的差异无统计学意义。研究发现两组 PV 患者的白细胞[（19．5士7．6）×109/L vs （7．1土1．1）×109/L，P ＝0．0005]、血小板计数[（498士172）×109/L vs （206士114）×109/L，P ＝0．0004]、ET 患者的血红蛋白[（152士18）g/L vs （112士11）g/L，P ＜O．OO01]、白细胞计数[（16．2士5．2）×109/L vs（9．3士3．4）×109/L，P ＜0．0001]以及 IMF 患者的白细胞[（15．1士3．8）×109/L vs （9．3土1．6）×109/L，P ＝0．0001]差异均有统计学意义。结论建立的多重 PCR方法可以同时检测5种 JAK2基因突变，可在临床上推广使用。与 JAK2突变阴性的 MPD 患者相比较，突变阳性的患者有更明显的骨髓增殖紊乱特征。
목적：건립다중등위기인특이성취합매련반응검측 JAK 2기인5충상견돌변적방법，병평개재3충상견적골수증식성질병중적림상의의。방법대149례 BCR-ABL 융합기인위음성적골수증식성질병（MPD）환자，포괄73례진성홍세포증다증（PV）、51례원발성혈소판증다증（ET）화25례원발성골수섬유화（IMF）환자；병결합림상급실험실기타검사자료，대기재해류질병진단화감별진단중적개치진행평고。20명급성백혈병환자화15명건강지원자작위대조조진행연구。소유병례적골수혹외주혈표본추제 DNA 후용건립적다중 PCR 방법진행평행검측。결과①149례 MPD 환자중공검출118례환자존재 JAK2기인돌변，총돌변솔위79．2％。기중 PV 환자양성68례，JAK2 V617F 돌변65례、JAK2 exon12돌변3례，양성솔93．2％（68/73）；ET 환자검측출양성35례，기중34례위 V617F 돌변，JAK2 exon12돌변1례，양성솔위68．6（35/51）；IMF 환자양성15례，균위 JAK2 V617F 돌변，양성솔위60％（15/25）。②JAK2돌변양성적 MPD 환자화돌변음성 MPD 적림상자료상비교，량조재환자성별，발병년빈방면적차이무통계학의의。연구발현량조 PV 환자적백세포[（19．5사7．6）×109/L vs （7．1토1．1）×109/L，P ＝0．0005]、혈소판계수[（498사172）×109/L vs （206사114）×109/L，P ＝0．0004]、ET 환자적혈홍단백[（152사18）g/L vs （112사11）g/L，P ＜O．OO01]、백세포계수[（16．2사5．2）×109/L vs（9．3사3．4）×109/L，P ＜0．0001]이급 IMF 환자적백세포[（15．1사3．8）×109/L vs （9．3토1．6）×109/L，P ＝0．0001]차이균유통계학의의。결론건립적다중 PCR방법가이동시검측5충 JAK2기인돌변，가재림상상추엄사용。여 JAK2돌변음성적 MPD 환자상비교，돌변양성적환자유경명현적골수증식문란특정。
Objective To establish the multiply PCR screening method of JAK2 and evaluate its clinical significance in patients with Myeloproliferative Disease (MPD).Methods 149 BCR-ABL negative myeloproliferative diseases (MPD)[73 polycythemia vera (PV)cases.51 primary Ethrombocytosis (ET)cases,25 myelofibrosis(lIMF)cases] were enrolled as study group;20 acute leukemia patients and 15 health individuals were enrolled as control group.Ge-nomic DNA from bone marrow or peripheral blood mononuclear cells was extracted and the JAK2 gene mutations of these subjects were detected by multiply AS-PCR.The clinical significance of JAK2 gene mutations and the potential usability of multiply AS-PCR were assessed,based on the clinical data of these patients.Results JAK2 mutations were detected in 118/149 BCR-ABL negative MPD patients,including 68/73 (93.2%,including 2 patients with JAK 2 ex-on12 mutation )PV patients,35/51 (68.6%,including 1 patients with JAK 2 exon12 mutation)ET patients and 15/25 (60.0 %)IMF patients.While the somatic JAK 2 mutations were not detected in any samples from control group.No difference in the clinical data in terms of age and sex was found between positive and negative MPD patients.However, there existed significant differences in leukocyte of PV patients [(19.5 ± 7.6)×109/L vs (7.1 ± 1.1)×109/L,P =0. 0005],platelet counts of PV patients [(498 ± 172)×109/L vs (206 ± 114)×109/L,P =0.0004],hemoglobin level of ET patients [(152 ± 18)g/L vs (112 ± 11)g/L,P <0.0001 ,leukocyte counts of ET patients [(16.2 ± 5.2)×109/L vs(9.3 ± 3.4)× 109/L,P <0.0001]and IMF patients [(15.1 ± 3.8)× 109/L vs (9.3 ± 1.6)× 109/L,P =0. 0001]between JAK 2 positíve and negative MPD patients.Conclusion Multiply As-PCR is one sensitive and reliable technique in detecting JAK 2 gene mutations.The clinical characters in JAK 2 positive MPD patients were more pro-gressing compared with those in JAK 2 negative patients.