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Annals of Oncology 13:781-788, 2002
© 2002 European Society for Medical Oncology

Original Paper

Quantitative measurement of BCR/abl transcripts using real-time polymerase chain reaction

W.-I. Lee1, H. Kantarjian2, A. Glassman3, M. Talpaz4 and M.-S. Lee1,+

1Molecular Diagnostics Laboratory, 3Cytogenetic Laboratory, Division of Pathology and Laboratory Medicine, 2Department of Leukemia, 4Department of Immunology and Biological Therapy, Division of Medicine, The University of Texas MD Anderson Cancer Center, Houston, USA

Received 2 May 2001; revised 13 November 2001; accepted 11 December 2001.

Background

Quantitative real-time polymerase chain reaction (Q-Rt-PCR) is a new tool in the detection and quantification of the BCR/abl fusion transcripts in chronic myelogenous leukemia (CML). This study investigates its specificity, sensitivity and potential clinical usefulness.

Patients and methods

Parallel analysis of Q-Rt-PCR and the conventional reverse transcription-mediated PCR (RT–PCR) were performed on 567 samples from 481 patients. Treatment response was monitored by Q-Rt-PCR at 6 and 12 months of 61 patients on STI-571 and 103 patients on interferon.

Results

The concordance rate between Q-Rt-PCR and RT–PCR was 96.3% (546/567), with 341 positives and 205 negatives. The positive equivalents ranged from 2 x 10–6 to 1.2 µg of K562 cell RNA. Karyotyping in 372 samples revealed excellent correlation with Q-Rt-PCR measurements (P <0.001). Setting residual BCR/abl <0.01 as an early goal of molecular response, we observed that STI-571 induced a better response than interferon: 49% (20 of 41 patients) versus 35% (15 of 62 patients) at 6 months (P = 0.025) and 52% (32 of 61 patients) versus 34% (35 of 103 patients) at 12 months (P = 0.01), respectively.

Conclusions

Q-Rt-PCR provides reliable measurements of BCR/abl fusion transcripts. It is potentially useful in assessing molecular residual disease after therapy.

Key words: chronic myelogenous leukemia, interferon, real-time polymerase chain reaction, STI-571


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