Open Access Research

Quantification of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL) using amplicon-fusion-site polymerase chain reaction (AFS-PCR)

Axel Weber12*, Sylvia Taube1, Udo zur Stadt3, Martin Horstmann4, Knut Krohn5, Jutta Bradtke6, Andrea Teigler-Schlegel6, Sabine Leiblein7 and Holger Christiansen1

Author Affiliations

1 Department of Pediatric Oncology, Hematology and Hemostaseology, Children’s Hospital, University of Leipzig, Leipzig, Germany

2 Department of Human Genetics, University Hospital Giessen & Marburg, Giessen, Germany

3 Center for Diagnostic, University Medical Center Hamburg Eppendorf, Hamburg, Germany

4 Research Institute Children’s Cancer Center Hamburg and Clinic of Pediatric Hematology and Oncology, University Medical Center Hamburg, Hamburg, Germany

5 IZKF Leipzig, University of Leipzig, Leipzig, Germany

6 Oncogenetic Laboratory - Department of Pediatric Hematology/Oncology, Justus Liebig-University, Giessen, Germany

7 Department of Hematology/Oncology, University of Leipzig, Leipzig, Germany

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Experimental Hematology & Oncology 2012, 1:33  doi:10.1186/2162-3619-1-33

Published: 9 November 2012


The amplification of putative oncogenes is a common finding within the genome of various cancer types. Identification and further targeting of specific junction sites within the sequence of genomic amplicons (amplicon fusion sites, AFS) by PCR (AFS-PCR) is suitable for quantification of minimal residual disease (MRD). This approach has recently been developed and described for MYCN amplified neuroblastomas. To compare AFS-PCR directly to routinely used MRD diagnostic strategies, we mapped the amplified genomic regions (ampGR) of an iAMP21-amplicon in high resolution of a patient with acute lymphoblastic leukemia (ALL). Successfully, we established AFS-PCR covering junction sites between ampGR within the iAMP21-amplicon. Quantification of MRD by AFS-PCR was directly comparable to IgH/TCR based real time quantitative PCR and fluorescence activated cell sorting (FACS) analysis in consecutive bone marrow (BM) specimens. Our data give an additional proof of concept of AFS-PCR for quantification of MRD. The method could be taken into account for ALL patients with genomic amplifications as alternative MRD diagnostic, if no or qualitatively poor Ig/TCR-PCRs are available.