Korean J Lab Med.  2003 Aug;23(4):221-228.

Monitoring of bcr-abl Fusion Transcript Levels by Quantitative Real-Time Polymerase Chain Reaction in Chronic Myeloid Leukemia after Bone Marrow Transplantation

• Choi SJ
• Chi HS
• Seo EJ
• Park CJ

Abstract

BACKGROUND
The bcr-abl gene rearrangement is a very important genetic marker in at least 95% of cases of chronic myeloid leukemia (CML). The technique that allows the detection of bcr-abl gene rearrangement seem to be the testing for the diagnoses and the detection of minimal residual disease (MRD) and may help in monitoring patients. Recent introduction of real-time quantitative PCR should eventually allow actual quantification of the target gene during amplification. In the present study, we investigated bcr-abl chimeric mRNA quantification in CML after bone marrow transplantation (BMT) by real-time RT-PCR and its clinical applicability to the decision of therapeutic intervention. METHODS: The subjects were a consecutive series of 9 CML patients with serial quantification of bcr-abl chimeric mRNA by real-time RT-PCR in peripheral blood or bone marrow aspirates. The sensitivity of real-time RT-PCR was compared to conventional RT-PCR and then the correlation with FISH results was analyzed. RESULTS: Seventeen bone marrow samples and 40 peripheral bloods were obtained from patients. The sensitivity of real-time RT-PCR was 10(-6) and positive signals were detected in negative cases by conventional RT-PCR. A significant correlation between the bcr-abl/G6PDH ratio by real-time RT-PCR and the proportion of positive cells for bcr-abl gene rearrangement by FISH was obtained (r=0.64). In the serial blood and bone marrow specimens, we found a progressive decrease in the bcr-abl/G6PDH ratio in all the patients. CONCLUSIONS: Our findings indicated that quantitative analysis of bcr-abl chimeric mRNA by real-time RT-PCR might be a useful tool for the monitoring of minimal residual disease in bcr-abl positive leukemic patients.

Keyword

Bcr-abl gene rearrangement; Chronic myeloid leukemia; Quantitative real-time RTPCR; Minimal residual disease; Bone marrow transplantation

MeSH Terms

Bone Marrow
Bone Marrow Transplantation*
Diagnosis
Gene Rearrangement
Genetic Markers
Humans
Leukemia, Myelogenous, Chronic, BCR-ABL Positive*
Neoplasm, Residual
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction*
RNA, Messenger
Genetic Markers
RNA, Messenger