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

Abstract

To determine the incidence of leukemiaspecific 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 welldefined 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. (Blood. 2001;97:805-808)

Acute myeloid leukaemia with 8p11 (MYST3) rearrangement: an integrated cytologic, cytogenetic and molecular study by the groupe francophone de cytogénétique hématologique

“…29 The gene in 19q13 is unknown. Neither inv(8)(p11q13) involving TIF2/NCOA2 (nuclear receptor coactivator 2), described in seven cases, 11,12,[30][31][32][33] Fwere found in our series.…”

Diagnosis of the Small Round Blue Cell Tumors Using Multiplex Polymerase Chain Reaction

“…Recently, multiple RT-PCR analyses have also been used as a screening tool for detection of genetic rearrangements, 9 for in vitro toxicology screening, 10 and for validation of gene sets obtained from global screens. 11 We verified the multiple RT-PCR results using the real-time RT-PCR method in a previous study.…”

A diagnostic biochip for the comprehensive analysis of MLL translocations in acute leukemia

“…However, additional analyses, such as split-out and modified post-PCR southern blot analysis are required for the detection and verification of an FG. [23][24][25][26] A recent publication describes a multiplex reaction followed by biochip detection to screen samples for the presence of translocations, but this technology is only applicable to well-characterized fusion events. 27 Moreover, the continual identification of new breakpoints, especially for MLL partner genes, as well as the technical complexity of this kind of assay, increase the risk of false negative and false positive results due to cross-contamination.…”