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Standardized next-generation sequencing of immunoglobulin and T-cell receptor gene recombinations for MRD marker identification in acute lymphoblastic leukaemia; a EuroClonality-NGS validation study

Monika Brüggemann 1, 2 Michaela Kotrova 3, 2 Henrik Knecht 4 Jack Bartram 2 Myriam Boudjogrha 5 Vojtech Bystry 6 Grazia Fazio 7 Eva Froňková 3 Mathieu Giraud 8 Andrea Grioni 7 Jeremy Hancock 9 Dietrich Herrmann 4 Cristina Jimenez 10 Adam Krejci 6 John Moppett 11 Tomas Reigl 6 Mikaël Salson 12, 8 Blanca Scheijen 13 Martin Schwarz 14, 4 Simona Songia 7 Michael Svaton 3 Jacques J. M. van Dongen 15 Patrick Villarese 16, 17 Stephanie Wakeman 9 Gary Wright 2 Giovanni Cazzaniga 7 Frédéric Davi 5 Ramón García-Sanz 18, 10 David Davi 5 Patricia Groenen 19 Michael Hummel 20 Elizabeth Macintyre 21 Kostas C Stamatopoulos 22 Christiane Pott 23, 4 Jan Trka 24, 3 Nikos Darzentas 6, 4 Anton Langerak 25 David Gonzalez 26 
12 BONSAI - Bioinformatics and Sequence Analysis
LIFL - Laboratoire d'Informatique Fondamentale de Lille, Inria Lille - Nord Europe
Abstract : Amplicon-based next-generation sequencing (NGS) of immunoglobulin (IG) and T-cell receptor (TR) gene rearrangements for clonality assessment, marker identification and quantification of minimal residual disease (MRD) in lymphoid neoplasms has been the focus of intense research, development and application. However, standardization and validation in a scientifically controlled multicentre setting is still lacking. Therefore, IG/TR assay development and design, including bioinformatics, was performed within the EuroClonality-NGS working group and validated for MRD marker identification in acute lymphoblastic leukaemia (ALL). Five EuroMRD ALL reference laboratories performed IG/TR NGS in 50 diagnostic ALL samples, and compared results with those generated through routine IG/TR Sanger sequencing. A central polytarget quality control (cPT-QC) was used to monitor primer performance, and a central in-tube quality control (cIT-QC) was spiked into each sample as a library-specific quality control and calibrator. NGS identified 259 (average 5.2/sample, range 0–14) clonal sequences vs. Sanger-sequencing 248 (average 5.0/sample, range 0–14). NGS primers covered possible IG/TR rearrangement types more completely compared with local multiplex PCR sets and enabled sequencing of bi-allelic rearrangements and weak PCR products. The cPT-QC showed high reproducibility across all laboratories. These validated and reproducible quality-controlled EuroClonality-NGS assays can be used for standardized NGS-based identification of IG/TR markers in lymphoid malignancies.
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https://hal.archives-ouvertes.fr/hal-02169053
Contributor : Mathieu Giraud Connect in order to contact the contributor
Submitted on : Sunday, June 30, 2019 - 7:43:15 AM
Last modification on : Friday, May 20, 2022 - 11:06:11 AM

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Monika Brüggemann, Michaela Kotrova, Henrik Knecht, Jack Bartram, Myriam Boudjogrha, et al.. Standardized next-generation sequencing of immunoglobulin and T-cell receptor gene recombinations for MRD marker identification in acute lymphoblastic leukaemia; a EuroClonality-NGS validation study. Leukemia, Nature Publishing Group: Open Access Hybrid Model Option B, 2019, ⟨10.1038/s41375-019-0496-7⟩. ⟨hal-02169053⟩

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