Monitoring Bcr-abl MRNA Levels In Chronic Myeloid Leukemia Patients By Real-time Quantitative RT-PCR

Objective:To establish real time quantitative RT-PCR(RQ–PCR) method for detection the mRNA levels of bcr-abl fusion gene in the patients with chronic myeloid leukemia (CML);and evaluate its clinical value in the minimal residual disease(MRD).Methods:To detect mRNA levels of bcr-abl fusion gene of 36 patients with CML by RQ -PCR and nested RT-PCR ,Ph chromosome using cytogenetic study.18 patients treated with allogene peripheral blood stem cell transplantation (Allo-PBSCT) were included.Results:1. RQ -PCR method could detect the bcr-abl molecules as low as 50 copies. 2.To detect bcr-abl fusion gene of 36 CML patients by RQ -PCR, the ration of positive were 94.4%(34/36) . The median expression level of bcr -abl fusion gene in 30 CML patients before therapy was 2.946(range: 0.025～10.255) .The expression levels of bcr-abl/GAPDH in 15 patients after 3 monthes ( 0.091 ) of Allo-PBSCT were significant lower than those before (1.317)transplantation(P<0.01)3. bcr-abl fusion gene was detected in 33 of 36 patients(91.6%) by the nested RT-PCR and was negative in 16 of 18 patients after 6 monthes of Allo-PBSCT.4.Philadelphia chromosome was detected in 34 of 36 patients(94.4%) by the cytogenetic study and was negative in 18 patients after 3 monthes of Allo-PBSCT.Conclusion:RQ -PCR is a sensitive and liable method to detect bcr- abl fusion gene .There is a difference in bcr- abl fusion gene expression levels among the newly diagnosed CML;This method can effective observe the change of bcr-abl mRNA before and post Allo-PBSCT in the Ph+ patient with CML .That is a liable method to monitor MRD of CML after allogeneic...