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Quality control and quantification in IG/TR next-generation sequencing marker identification: protocols and bioinformatic functionalities by EuroClonality-NGS

AuthorsKnecht, Henrik; Reigl, Tomas; Kotrová, M.; Appelt, Franziska; Stewart, Peter; Bystry, Vojtech; Krejci, Adam; Grioni, Andrea; Pal, Karol; Stranska, Kamila; Plevova, Karla; Rijntjes, Jos; Songia, Simona; Svatoň, Michael; Froňková, Eva; Bartram, Jack; Scheijen, Blanca; Herrmann, Dietrich; García-Sanz, Ramón; Hancock, Jeremy; Moppett, John; Dongen, J. J. M. van; Cazzaniga, Giovanni; Davi, Frédéric; Groenen, Patricia J. T. A.; Hummel, Michael; Macintyre, Elizabeth A.; Stamatopoulos, Kostas; Trka, Jan; Langerak, Anton W.; González, David; Pott, Christiane; Brüggemann, Monika; Darzentas, Nikos
KeywordsCancer genetics
Genetics research
Issue Date2019
PublisherSpringer Nature
CitationLeukemia 33: 2254–2265 (2019)
AbstractAssessment of clonality, marker identification and measurement of minimal residual disease (MRD) of immunoglobulin (IG) and T cell receptor (TR) gene rearrangements in lymphoid neoplasms using next-generation sequencing (NGS) is currently under intensive development for use in clinical diagnostics. So far, however, there is a lack of suitable quality control (QC) options with regard to standardisation and quality metrics to ensure robust clinical application of such approaches. The EuroClonality-NGS Working Group has therefore established two types of QCs to accompany the NGS-based IG/TR assays. First, a central polytarget QC (cPT-QC) is used to monitor the primer performance of each of the EuroClonality multiplex NGS assays; second, a standardised human cell line-based DNA control is spiked into each patient DNA sample to work as a central in-tube QC and calibrator for MRD quantification (cIT-QC). Having integrated those two reference standards in the ARResT/Interrogate bioinformatic platform, EuroClonality-NGS provides a complete protocol for standardised IG/TR gene rearrangement analysis by NGS with high reproducibility, accuracy and precision for valid marker identification and quantification in diagnostics of lymphoid malignancies.
Publisher version (URL)http://dx.doi.org/10.1038/s41375-019-0499-4
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