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Multiplex Reverse Transcription-Polymerase Chain
Reaction for Simultaneous Screening of 29 Translocations
and Chromosomal Aberrations in Acute Leukemia
Niels Pallisgaard,
Peter Hokland,
Dorthe C. Riishøj,
Bent Pedersen, and
Poul Jørgensen
From the Department of Molecular and Structural Biology, Aarhus
University, Aarhus, Denmark; the Department of Hematology and Medicine,
Aarhus University Hospital, Aarhus, Denmark; and the Department of
Cytogenetics, Danish Cancer Society, Aarhus, Denmark.
We have developed a multiplex reverse transcription-polymerase chain
reaction (RT-PCR) reaction, which enables us to detect 29 translocations/chromosomal aberrations in patients with acute lymphoid
leukemia (ALL) and acute myeloid leukemia (AML). Through the
construction and optimization of specific primers for each translocation, we have been able to reduce the set-up to 8 parallel multiplex PCR reactions, thus greatly decreasing the amount of work and
reagents. We show the value of our set-up in a retrospective analysis
on cryopreserved material from 102 AML and 62 ALL patients. The
multiplex RT-PCR detected a hybrid mRNA resulting from a structural chromosomal aberration in 45 of 102 (44%) of the AML and in 28 of 62 (45%) of the pediatric ALL cases. Importantly, in 33% of AML and in
47% of the ALL cases with cytogenetic data, submicroscopic chromosomal
aberrations or masked translocations were shown that were not detected
in the cytogenetic analysis either for structural reasons or because of
an insufficient number of metaphases obtained. This multiplex RT-PCR
system, which can handle up to 10 patients with a response time of 2 working days, is thus an important tool that complements cytogenetic
analysis in the up-front screening of acute leukemia patients and
should provide a rapid and efficient characterization of leukemia
cells, even in situations with sparse patient material.
Blood, Vol. 92 No. 2 (July 15), 1998:
pp. 574-588
© 1998 by the American Society of Hematology.
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