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


ABSTRACT: Assessment 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.

SUBMITTER: Knecht H 

PROVIDER: S-EPMC6756032 | biostudies-literature | 2019 Sep

REPOSITORIES: biostudies-literature

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

Knecht Henrik H   Reigl Tomas T   Kotrová Michaela M   Appelt Franziska F   Stewart Peter P   Bystry Vojtech V   Krejci Adam A   Grioni Andrea A   Pal Karol K   Stranska Kamila K   Plevova Karla K   Rijntjes Jos J   Songia Simona S   Svatoň Michael M   Froňková Eva E   Bartram Jack J   Scheijen Blanca B   Herrmann Dietrich D   García-Sanz Ramón R   Hancock Jeremy J   Moppett John J   van Dongen Jacques J M JJM   Cazzaniga Giovanni G   Davi Frédéric F   Groenen Patricia J T A PJTA   Hummel Michael M   Macintyre Elizabeth A EA   Stamatopoulos Kostas K   Trka Jan J   Langerak Anton W AW   Gonzalez David D   Pott Christiane C   Brüggemann Monika M   Darzentas Nikos N  

Leukemia 20190621 9


Assessment 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  ...[more]

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