TY  - JOUR
A1  - Jann, Johann-Christoph
A1  - Nowak, Daniel
A1  - Nolte, Florian
A1  - Fey, Stephanie
A1  - Nowak, Verena
A1  - Obländer, Julia
A1  - Pressler, Jovita
A1  - Palme, Iris
A1  - Xanthopoulos, Christina
A1  - Fabarius, Alice
A1  - Platzbecker, Uwe
A1  - Giagounidis, Aristoteles
A1  - Götze, Katharina
A1  - Letsch, Anne
A1  - Haase, Detlef
A1  - Schlenk, Richard Friedrich
A1  - Bug, Gesine
A1  - Lübbert, Michael
A1  - Ganser, Arnold
A1  - Germing, Ulrich
A1  - Haferlach, Claudia
A1  - Hofmann, Wolf-Karsten
A1  - Mossner, Maximilian
T1  - Accurate quantification of chromosomal lesions via short tandem repeat analysis using minimal amounts of DNA
T2  - Journal of medical genetics
N2  - Background: Cytogenetic aberrations such as deletion of chromosome 5q (del(5q)) represent key elements in routine clinical diagnostics of haematological malignancies. Currently established methods such as metaphase cytogenetics, FISH or array-based approaches have limitations due to their dependency on viable cells, high costs or semi-quantitative nature. Importantly, they cannot be used on low abundance DNA. We therefore aimed to establish a robust and quantitative technique that overcomes these shortcomings.
Methods: For precise determination of del(5q) cell fractions, we developed an inexpensive multiplex-PCR assay requiring only nanograms of DNA that simultaneously measures allelic imbalances of 12 independent short tandem repeat markers.
Results: Application of this method to n=1142 samples from n=260 individuals revealed strong intermarker concordance (R²=0.77–0.97) and reproducibility (mean SD: 1.7%). Notably, the assay showed accurate quantification via standard curve assessment (R²>0.99) and high concordance with paired FISH measurements (R²=0.92) even with subnanogram amounts of DNA. Moreover, cytogenetic response was reliably confirmed in del(5q) patients with myelodysplastic syndromes treated with lenalidomide. While the assay demonstrated good diagnostic accuracy in receiver operating characteristic analysis (area under the curve: 0.97), we further observed robust correlation between bone marrow and peripheral blood samples (R²=0.79), suggesting its potential suitability for less-invasive clonal monitoring.
Conclusions: In conclusion, we present an adaptable tool for quantification of chromosomal aberrations, particularly in problematic samples, which should be easily applicable to further tumour entities.
KW  - Chromosomal deletion
KW  - PCR
KW  - deletion 5q
KW  - myelodysplastic syndrome
KW  - short tandem repeats
Y1  - 2017
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/46600
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-466009
SN  - 1468-6244
SN  - 0022-2593
N1  - Copyright information: © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted. This is an Open Access article distributed in accordance with the terms of the Creative Commons Attribution (CC BY 4.0) license, which permits others to distribute, remix, adapt and build upon this work, for commercial use, provided the original work is properly cited. See: http://creativecommons.org/licenses/by/4.0/
VL  - 54
IS  - 9
SP  - 640
EP  - 650
PB  - BMJ Publishing Group
CY  - London
ER  -