Detection and quantification ofCBFB/MYH11 fusion transcripts in patients with inv(16)-positive acute myeloblastic leukemia by real-time RT-PCR

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Abstract
We used a newly established real‐time RT‐PCR assay for the quantification of the leukemia‐specific CBFB/MYH11 transcripts in inv(16)‐positive acute myeloblastic leukemia. CBFB/MYH11 could be quantified over a five log range, with a detection limit of 10 molecules of a CBFB/MYH11 plasmid and a 1:105 dilution of RNA of the inv(16)‐positive ME‐1 cell line, respectively. The fusion transcripts were also quantified in 19 patients with acute myeloblastic leukemia and an inv(16) at initial diagnosis. The expression of CBFB/MYH11 varied over a two log range without correlation to clinical response or relapse rate. In nine patients, CBFB/MYH11 was also quantified during/after chemotherapy and autologous or allogeneic stem cell transplantation. All of these patients showed a similar decline of CBFB/MYH11 after intensive therapy. Six of these patients are in complete remission with a stable low‐level or absent CBFB/MYH11 expression. Three patients relapsed, and their CBFB/MYH11 transcripts rose again to pretreatment levels. In two patients, this increase in CBFB/MYH11 could be detected by real‐time PCR before hematological relapse. These data indicate that real‐time RT‐PCR can be used for the sensitive detection and quantification of CBFB/MYH11 transcripts in the follow‐up of patients with inv(16)‐positive AML.

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