摘要目的 观察实时定量聚合酶链反应(PCR)检测慢性骨髓增殖性疾病(CMPD)中JAK2基因V617F点突变情况.方法 采用实时定量PCR法检测56例CMPD患者JAK2 V617F基因突变类型及突变转录水平.其中,真性红细胞增多症(PV)26例、原发性血小板增多症(ET)24例、原发性骨髓纤维化(CIMF)5例、高嗜酸粒细胞综合征(HES)1例.结果 56例CMPD患者JAK2 V617F基因突变率为51.79%(29/56),PV 65.38%(17/26)、ET 37.50%(9/24)、IMF60.00%(3/5);杂合性突变率为41.07%(23/56),包括PV 53.85%(14/26)、ET 29.17%(7/24)、CIMF 40.0%(2/5);纯合性突变率为10.71%(6/56),包括PV 11.54%(3/26)、ET 8.33%(2/24)、CIMF 20.00%(1/5);1例HES患者为野生型.PV、ET和CIMF突变患者拷贝数分别为2-14×102～1.5×1 07、9.80×102～4.4×107和4.10×103～3.70×106.结论 实时定量PCR检测JAK2V617F基因简便、快捷,并且易于定量,检测阳性率与国外报道相似,适合临床用于CMPD的诊断和疗效评估.

abstractsObjective To investigate the novel V617F point mutation of JAK2 gene by real-time PGR in the patients with chronic myeloproliferative disease (CMPD) and evaluate its clinical significance. Methods Genomic DNA from bone marrow or peripheral blood mononuclear cells was extracted from 56 patients with CMPD, including 26 cases of polycythemia vera (PV), 24 cases of essential thrombocythemia (ET), 5 cases of chronic idiopathicmyelofibrosis (CIMF) and 1 case of high eosinophilic syndrome (HES). The exon 14 of JAK2 gene which harbourd V617F mutation were screened by real-time PGR. Results JAK2 V617F mutation was measured in 29 of the 56 patients with CMPD. The prevalence of mutation was 65.38 %(17/26) in PV,37.50 % in ET and 60.00 %(3/5) in CIMF. The proportion of mutation in PV, ET and CIMF are respectively 53.85 %(14/26), 29.17 %(7/24), 40.0 %(2/5) in heterozygotes and 11.54 %(3/26), 8.33 %(2/24), 20.00 %(1/5)in homozygotes. JAK2 mutation was negative in one patient with HES. JAK2 V617F allele burden in PV, ET and CIMF are respectively 2.14×102-1.5×107, 9.80×102-4.4×107 and 4.10×103-3.70×106 copies. Conclusion Real-time PCR is a useful tool for pre cisely assess the grade of mutant allele burden as well as to screen JAK2V617F mutation simultaneously, which is simple and convenient to carry out in clinical laboratories for diagnosis and further evaluations of minimal residual disease in CMPD patients.