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  • Letter to the Editor
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Rational use of the EAC real-time quantitative PCR protocol in chronic myelogenous leukemia: report of three false-negative cases at diagnosis

Leukemia volume 20, pages 886–888 (2006)Cite this article

This letter is addressed to the paper by Gabert et al.,1 in which the Europe Against Cancer (EAC) program has proposed a standardized real-time quantitative polymerase chain reaction (RQ-PCR) protocol designed for studies of minimal residual disease (MRD) in fusion transcript bearing leukemia. Although this protocol has been established in the frame of molecular follow-up, its very practical use would prompt molecular labs to choose it as tool for diagnostic purposes, at least for BCR-ABL screening in chronic myelogenous leukemia (CML). This is an inappropriate approach as false-negative results at diagnosis of a CML may be encountered with the use of this protocol. Indeed, we report hereby three Philadelphia (Ph+) CML patients with a major breakpoint cluster region (M-bcr) rearrangement detectable by fluorescent in situ hybridization (FISH) and classical qualitative RT-PCR but not by RQ-PCR with the use of the standardized EAC protocol.

We are thus reporting three Ph+ CML cases with M-BCR-ABL rearrangement detectable by FISH and qualitative RT-PCR, but not by RQ-PCR (Taqman technology) using the EAC set of primers and probe targeting both e13a2 and e14a2 fusion transcripts. The possibility of an e19a2 BCR-ABL junction (p230Bcr-Abl) or rare atypical BCR-ABL transcripts involving various BCR exons2, 3 was ruled out by qualitative PCR and sequencing. Also, the FISH profile observed (1F1R1G and one extra signal) was not in keeping with an e1a2 junction, which leads to the production of p190Bcr-Abl and occurs very rarely in CML.2 In our three cases, the t(9;22) generated classical e13a2 transcripts, but the failure to identify any BCR-ABL transcript by RQ-PCR was owing to sequence rearrangements targeting the forward BCR primer hybridization site located at the border of the BCR-ABL junction.

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References

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Authors and Affiliations

  1. Department of Medical Genetics, University Hospital Erasme, Brussels, Belgium

    B Dessars, H El Housni & P Heimann

  2. Department of Human Genetics, Molecular Hematology Unit, Pathology Institute, CHU Sart-Tilman, Liege, Belgium

    F Lambert

  3. Department of Hematology, University Hospital Erasme, Brussels, Belgium

    A Kentos

  4. Laboratory of Cytogenetics, Jules Bordet Institute, Brussels, Belgium

    P Heimann

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  1. B Dessars
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  2. H El Housni
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Correspondence to P Heimann.

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Dessars, B., El Housni, H., Lambert, F. et al. Rational use of the EAC real-time quantitative PCR protocol in chronic myelogenous leukemia: report of three false-negative cases at diagnosis. Leukemia 20, 886–888 (2006). https://doi.org/10.1038/sj.leu.2404174

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