Portal de Periódicos da CAPES

Multiplex reverse transcriptase–polymerase chain reaction screening in childhood acute myeloblastic leukemia

2001; Elsevier BV; Volume: 97; Issue: 3 Linguagem: Inglês

10.1182/blood.v97.3.805

1528-0020

Sabine Strehl, Margit König, Georg Mann, Oskar A. Haas,

Advanced biosensing and bioanalysis techniques

Abstract To determine the incidence of leukemia-specific rearrangements, 60 cases of childhood acute myeloblastic leukemia and transient myeloproliferative disorder were screened with a novel multiplex reverse transcriptase–polymerase chain reaction (RT-PCR) assay, and the results were correlated with the cytogenetic findings. The RT-PCR assay detects 28 different fusion genes and more than 80 different fusion transcript variants. RNA was isolated from methanol/acetic acid–fixed cells that had been routinely prepared for cytogenetic analysis. Nine different fusion transcripts were found in 40% of the cases, whereas 78.3% of the cases had abnormal karyotypes. Two cases with a t(6;11) and an MLL/AF6 gene fusion were missed cytogenetically. Conversely, cytogenetic analysis revealed 10 other well-defined chromosome rearrangements. Although cytogenetic analysis reveals a much broader range of abnormalities, multiplex RT-PCR serves as quality control and provides the essential information for minimal residual disease studies. Moreover, discrepant findings lead to the detection of new rearrangements on the molecular genetic level.