TY - JOUR
T1 - Next-Generation Sequencing–Based Clonality Assessment of Ig Gene Rearrangements
T2 - A Multicenter Validation Study by EuroClonality-NGS
AU - EuroClonality-NGS Working Group
AU - van den Brand, Michiel
AU - Rijntjes, Jos
AU - Möbs, Markus
AU - Steinhilber, Julia
AU - van der Klift, Michèle Y.
AU - Heezen, Kim C.
AU - Kroeze, Leonie I.
AU - Reigl, Tomas
AU - Porc, Jakub
AU - Darzentas, Nikos
AU - Luijks, Jeroen A.C.W.
AU - Scheijen, Blanca
AU - Davi, Frédéric
AU - ElDaly, Hesham
AU - Liu, Hongxiang
AU - Anagnostopoulos, Ioannis
AU - Hummel, Michael
AU - Fend, Falko
AU - Langerak, Anton W.
AU - Groenen, Patricia J.T.A.
N1 - Funding Information:
Supported by the Dutch Health Insurers' Innovation Fund project number 17-179; and the revenues of the previously obtained patent ( PCT/NL2003/000690 ), which is collectively owned by the EuroClonality/BIOMED-2 Consortium and licensed to Invivoscribe.
Publisher Copyright:
© 2021 Association for Molecular Pathology and American Society for Investigative Pathology
PY - 2021/9/1
Y1 - 2021/9/1
N2 - Ig gene (IG) clonality analysis has an important role in the distinction of benign and malignant B-cell lymphoid proliferations and is mostly performed with the conventional EuroClonality/BIOMED-2 multiplex PCR protocol and GeneScan fragment size analysis. Recently, the EuroClonality-NGS Working Group developed a method for next-generation sequencing (NGS)–based IG clonality analysis. Herein, we report the results of an international multicenter biological validation of this novel method compared with the gold standard EuroClonality/BIOMED-2 protocol, based on 209 specimens of reactive and neoplastic lymphoproliferations. NGS-based IG clonality analysis showed a high interlaboratory concordance (99%) and high concordance with conventional clonality analysis (98%) for the molecular conclusion. Detailed analysis of the individual IG heavy chain and kappa light chain targets showed that NGS-based clonality analysis was more often able to detect a clonal rearrangement or yield an interpretable result. NGS-based and conventional clonality analysis detected a clone in 96% and 95% of B-cell neoplasms, respectively, and all but one of the reactive cases were scored polyclonal. We conclude that NGS-based IG clonality analysis performs comparable to conventional clonality analysis. We provide critical parameters for interpretation and discuss a first step toward a quantitative scoring approach for NGS clonality results. Considering the advantages of NGS-based clonality analysis, including its high sensitivity and possibilities for accurate clonal comparison, this supports implementation in diagnostic practice.
AB - Ig gene (IG) clonality analysis has an important role in the distinction of benign and malignant B-cell lymphoid proliferations and is mostly performed with the conventional EuroClonality/BIOMED-2 multiplex PCR protocol and GeneScan fragment size analysis. Recently, the EuroClonality-NGS Working Group developed a method for next-generation sequencing (NGS)–based IG clonality analysis. Herein, we report the results of an international multicenter biological validation of this novel method compared with the gold standard EuroClonality/BIOMED-2 protocol, based on 209 specimens of reactive and neoplastic lymphoproliferations. NGS-based IG clonality analysis showed a high interlaboratory concordance (99%) and high concordance with conventional clonality analysis (98%) for the molecular conclusion. Detailed analysis of the individual IG heavy chain and kappa light chain targets showed that NGS-based clonality analysis was more often able to detect a clonal rearrangement or yield an interpretable result. NGS-based and conventional clonality analysis detected a clone in 96% and 95% of B-cell neoplasms, respectively, and all but one of the reactive cases were scored polyclonal. We conclude that NGS-based IG clonality analysis performs comparable to conventional clonality analysis. We provide critical parameters for interpretation and discuss a first step toward a quantitative scoring approach for NGS clonality results. Considering the advantages of NGS-based clonality analysis, including its high sensitivity and possibilities for accurate clonal comparison, this supports implementation in diagnostic practice.
UR - http://www.scopus.com/inward/record.url?scp=85113384817&partnerID=8YFLogxK
U2 - 10.1016/j.jmoldx.2021.06.005
DO - 10.1016/j.jmoldx.2021.06.005
M3 - Article
C2 - 34186174
AN - SCOPUS:85113384817
SN - 1525-1578
VL - 23
SP - 1105
EP - 1115
JO - Journal of Molecular Diagnostics
JF - Journal of Molecular Diagnostics
IS - 9
ER -