实时定量聚合酶链反应与巢式聚合酶链反应在289例白血病融合基因检测中的比较

摘要

Objective To explore the differences of real-time quantitative polymerase chain reaction (RT-PCR) and nested polymerase chain reaction (nPCR) in the detection of fusion genes of BCR/ABL,PMWRARa and AML1/ET0 in leukemia. Methods RT-PCR and nPCR were applied for detection of the fusion genes BCR/ABL,PML/RARa and AML1/ET0 in 289 patients with leukemia,the results combined with cytomophology and clinical outcomes of patients were analyzed. Results The positive rates of fusion gene of BCR/ABL detected by RT-PCR and nPCR in 46 cases of newly diagnosed chronic myelocytic leukemia (CML) were 95.7% and 93.5% respectively (P = 0.997),while the ratios were 78.4% and 58.1% (P < 0.05) in 69 cases of complete remission (CR). By the same detections,the positive rates of fusion gene of PML/RARa in 32 cases of newly diagnosed acute promyelocytic leukemia (APL) were 93.8% and 90.6% respectively (P =0.996),whae the ratios were 12.3% and 7.0% (P<0.05) in 114 cases of CR. The positive rates of fusion gene of AML1/ETO in 12 cases of newly diagnosed acute myeloblastic leukemia M2 (ANLL-M2) were 50.0% and 50.0% respectively (P =1.0),while the ratios were 50.0% and 12.5% (P < 0.05) in 16 cases of CR. Conclusion RT-PCR is more sensitive and accurate than nPCR for detecting fusion genes of leukemia. It can improve the detection of minimal residual disease, and provide more effective molecular biological basis for diagnosis and treatment in leukemia.%目的 比较实时定量聚合酶链反应(RT-PCR)与巢式聚合酶链反应(nPCR)在白血病融合基因BCR/ABL、PML/RARa及AML1/ETO检测中的结果.方法 分别应用RT-PCR与nPCR对289例急慢性白血病初诊和完全缓解的患者BCR/ABL、PML/RARa及AML1/ETO融合基因进行检测,结合患者骨髓细胞形态学及临床转归予以分析.结果 46例初诊慢性粒细胞白血病(CML)患者中,RT-PCR与nPCR检测BCR/ABL融合基因阳性率分别为95.7％和93.5％(P=0.997)；69例完全缓解CML患者中,RT-PCR与nPCR检测阳性率分别为78.4％和58.1％(P＜0.05).32例初诊急性早幼粒细胞白血病(APL)患者中,RT-PCR与nPCR检测PML/RARa融合基因阳性率分别为93.8％和90.6％(P=0.996)；114例完全缓解APL患者中,RT-PCR与nPCR检测阳性率分别为12.3％和7.0％(P＜0.05).12例初诊急性粒细胞白血病M2(ANLL-M2)患者中,RT-PCR与nPCR检测AML1/ETO融合基因阳性率分别为50.0％和50.0％(P=1.0)；16例完全缓解ANLL-M2患者中,RT-PCR与nPCR检测阳性率分别为50.0％和12.5％(P＜0.05).结论 RT-PCR较nPCR更为敏感而准确,应用RT-PCR可以提高微小残留病的检出率,为临床诊断和治疗提供更为有效的分子生物学依据.