白血病bcr/abl mRNA的荧光定量RTP-CR检测

REAL-TIME QUANTITATION OF bcr/abl TRANSCRIPTS IN LEUKEMIA WITH QUANTITATIVE RT-PCR

①目的 探讨bcr/abl基因表达水平与白血病诊断、预后的关系及在微小残留病检测中的意义。②方法 荧光定量RT PCR法 (FQ RT PCR)测定 34例不同类型及处于不同疾病阶段的白血病病人bcr/abl基因的表达。③结果 对照组bcr/abl基因表达均为阴性。慢性粒细胞白血病 (CML)病人bcr/abl基因表达阳性率为 89.5 % (1 7/ 1 9) ,经治疗达临床缓解者 8例 ,bcr/abl基因表达均值为 (2 .4 3± 0 .6 7)× 1 0 5基因拷贝 /L ;初诊CML未经治疗及治疗无效并发生急变或死亡者bcr/abl基因表达均值为 (2 .6 0± 0 .90 )× 1 0 7基因拷贝 /L ,两者有极显著性差异(t=3.4 3,P <0 .0 1 )。随访 4例CML ,2例治疗缓解 ,bcr/abl转录减少 ,无阴转 ;持续增高 2例中 ,1例急变 ,1例死亡。急性淋巴细胞白血病 (ALL)病人bcr/abl表达阳性率为 1 1 .8% (2 / 1 5 ) ,均复发 ,1例死亡 ,bcr/abl基因转录复发前表达明显增高。④结论 FQ RT PCR实验方法灵敏、快速、特异性好、结果准确 ;bcr/abl基因表达水平的定量观察及动态监测 ,有助于白血病的临床诊断与分型 。

Objective To assess the relationship between expression of bcr/abl mRNA and the diagnosis as well as prognosis of leukemia, and the significance in the detection of minimal residual disease (MRD). Methods Expression of bcr/abl gene was measured with fluorogenic probe quantitative RT-PCR(FQ-RT-PCR) in 34 cases of leukemia at different stages. Results No bcr/abl gene was revealed in the controls, while in the chronic myelogenous leukemia(CML) group the positive rate was 89.5%(17/19). Eight patients were relieved after α-interferon therapy, with an average bcr/abl gene expression of (2.43±0.67)×10 5 copies/L, while in those untreated the average expression was (2.60±0.90)×10 7 copies/L. The difference between two groups was significant ( t=3.43, P <0.01). Follow-up data were obtained in six bcr/abl positive cases, with reduced bcr/abl gene transcription but no negative transformation in two cases, and increased transcription in the other two. Conclusion Being sensitive, specific, reliable and accurate, FQ-RT-PCR assay may detect dynamic evolution of bcr/abl transcripts, and help to make the diagnosis and typing of leukemia, and to predict the existence of MRD.