Real-time quantification of different types of BcrAbl transcript in chronic myeloid leukemia

Barbara B. Giannini, Vittorio V. Montefusco, Claudia C. Zardini, Nicoletta N. Testoni, Gianantonio G. Rosti, Carolina C. Terragna, Silvia S. Buonamici, Emanuela E. Ottaviani, Simona S. Bassi, Antonio A. De Vivo, Michele M. Baccarani, Giuseppe G. Saglio, Sante S. Tura

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Abstract

The most common translocation in chronic myeloid leukemia (CML), t(9;22)(q34;q22), produces the BCR/ABL fusion gene. We set up and evaluated a rapid and reliable real-time RT-PCR approach using TaqMan technology for detection and quantification of bcr-abl transcripts in CML patients at diagnosis and during therapy. A pair of primers and probe complementary to ABL exon 2 were designed, enabling detection of the most frequent bcr-abl transcripts, and of normal ABL-Ia as an internal control. Conditions were established to amplify less than 1-10 target molecules/reaction and detect one CML cell in 106 cells from healthy donors. To determine the utility of the assay, we quantified the bcr-abl/ABL-Ia ratio in 47 bone marrow from 33 CML patients at diagnosis and 14 in cytogenetic remission (CR). In 14 cases, this ratio was compared with results obtained by a competitive-quantitative RT-PCR/capillary electrophoresis method from contemporary specimens. By real-time RT-PCR, the median value of bcr-abl/ABL-Ia ratio at diagnosis was 15.334 (range 28.810-3.361) and fell to 0.0012 (range 0.003-1.34) in CR. The real-time bcr-abl/ ABL-Ia ratios correlated with those obtained by competitive RT-PCR (p

Original languageEnglish
JournalBlood
Volume96
Issue number11 PART II
Publication statusPublished - 2000

ASJC Scopus subject areas

  • Hematology

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    Giannini, B. B., Montefusco, V. V., Zardini, C. C., Testoni, N. N., Rosti, G. G., Terragna, C. C., Buonamici, S. S., Ottaviani, E. E., Bassi, S. S., De Vivo, A. A., Baccarani, M. M., Saglio, G. G., & Tura, S. S. (2000). Real-time quantification of different types of BcrAbl transcript in chronic myeloid leukemia. Blood, 96(11 PART II).