Genomic inverse PCR for exploration of ligated breakpoints (GIPFEL), a new method to detect translocations in leukemia

Füller E, Schaefer D, Fischer U, Krell PFI, Stanulla M, Borkhardt A, Slany R (2014)

Publication Status: Published

Publication Type: Journal article, Original article

Publication year: 2014

Journal

PLoS ONE Public Library of Science

Book Volume: 9

Article Number: e104419

Journal Issue: 8

DOI: 10.1371/journal.pone.0104419

Abstract

Here we present a novel method "Genomic inverse PCR for exploration of ligated breakpoints" (GIPFEL) that allows the sensitive detection of recurrent chromosomal translocations. This technique utilizes limited amounts of DNA as starting material and relies on PCR based quantification of unique DNA sequences that are created by circular ligation of restricted genomic DNA from translocation bearing cells. Because the complete potential breakpoint region is interrogated, a prior knowledge of the individual, specific interchromosomal fusion site is not required. We validated GIPFEL for the five most common gene fusions associated with childhood leukemia (MLL-AF4, MLL-AF9, MLL-ENL, ETV6-RUNX1, and TCF3-PBX1). A workflow of restriction digest, purification, ligation, removal of linear fragments and precipitation enriching for circular DNA was developed. GIPFEL allowed detection of translocation specific signature sequences down to a 10^-4 dilution which is close to the theoretical limit. In a blinded proof-of-principle study utilizing DNA from cell lines and 144 children with B-precursor-ALL associated translocations this method was 100% specific with no false positive results. Sensitivity was 83%, 65%, and 24% for t(4;11), t(9;11) and t(11;19) respectively. Translocation t(12;21) was correctly detected in 64% and t(1;19) in 39% of the cases. In contrast to other methods, the characteristics of GIPFEL make it particularly attractive for prospective studies. © 2014 Fueller et al.

Authors with CRIS profile

Robert Slany Lehrstuhl für Genetik

Involved external institutions

Heinrich-Heine-Universität Düsseldorf

Germany (DE) Medizinische Hochschule Hannover (MHH) / Hannover Medical School

Germany (DE)

How to cite

APA:

Füller, E., Schaefer, D., Fischer, U., Krell, P.F.I., Stanulla, M., Borkhardt, A., & Slany, R. (2014). Genomic inverse PCR for exploration of ligated breakpoints (GIPFEL), a new method to detect translocations in leukemia. PLoS ONE, 9(8). https://doi.org/10.1371/journal.pone.0104419

MLA:

Füller, Elisa, et al. "Genomic inverse PCR for exploration of ligated breakpoints (GIPFEL), a new method to detect translocations in leukemia." PLoS ONE 9.8 (2014).

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