TY - JOUR
T1 - Real-time quantification of different types of BcrAbl transcript in chronic myeloid leukemia
AU - Giannini, Barbara B.
AU - Montefusco, Vittorio V.
AU - Zardini, Claudia C.
AU - Testoni, Nicoletta N.
AU - Rosti, Gianantonio G.
AU - Terragna, Carolina C.
AU - Buonamici, Silvia S.
AU - Ottaviani, Emanuela E.
AU - Bassi, Simona S.
AU - De Vivo, Antonio A.
AU - Baccarani, Michele M.
AU - Saglio, Giuseppe G.
AU - Tura, Sante S.
PY - 2000
Y1 - 2000
N2 - 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
AB - 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
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M3 - Article
AN - SCOPUS:33748549605
VL - 96
JO - Blood
JF - Blood
SN - 0006-4971
IS - 11 PART II
ER -