Detection of bcr-abl Fusion Gene in Chronic Myelocytic Leukemia Patients by Reverse Transcriptase Polymerse Chain Reation / 医学研究杂志

ABSTRACT

Objective To increase the sensitivity of residual leukemic cells detectin in chronic myelocytic leukemia(CML) patients with RT-PCR,the optional annealing temperature and PCR cycles were studied to confirm bcr-abl fused gene types,and bcr-abl mRNA transcripts were monitored by RQ-PCR to study the relation with CML at different phases. Methods Through changing the PCR conditions, the annealing temperature was measured from 55℃ to 60℃, and the number of reaction cycles was increased from 30 to 45.All 22 samples were examined, and bcr-abl mRNA transcripts were quantified by RQ-PCR kit. Results Bcr-abl fused gene types were found in 22 samples,of all 9 cases were b_2a_2 type, 13 cases were b_3a_2.When the annealing temperature was set for 58℃ and the number of reation cycles was set for 45,10~3 copies/ul standard samples was detected.18 samples were positive tested by RQ-PCR kit,and the value was between 10~2 to 10~6 copies/g.There were significant differce between the results of chronic phase samples and those of accelerated phase. Conclusions The RT-PCR is a reliable,sensitive and reproducible method of monitoring CML patients.The real-time RT-PCR is useful in evaluating leukemic burden,assessing response to treatment and predicating the prognosis of the disease.