Analysis and interpretation of Ig and TCR gene rearrangements in the conventional, low-throughput way have their limitations in terms of resolution, coverage, and biases. With the advent of high-throughput, next-generation sequencing (NGS) technologies, a deeper analysis of Ig and/or TCR (IG/TR) gene rearrangements is now within reach, which impacts on all main applications of IG/TR immunogenetic analysis. To bridge the generation gap from low- to high-throughput analysis, the EuroClonality-NGS Consortium has been formed, with the main objectives to develop, standardize, and validate the entire workflow of IG/TR NGS assays for 1) clonality assessment, 2) minimal residual disease detection, and 3) repertoire analysis. This concerns the preanalytical (sample preparation, target choice), analytical (amplification, NGS), and postanalytical (immunoinformatics) phases. Here we critically discuss pitfalls and challenges of IG/TR NGS methodology and its applications in hemato-oncology and immunology.
↵1 All listed authors are members of the EuroClonality-NGS Consortium and have written this article on behalf of the entire consortium.
See related articles in this issue: IJspeert et al. (J. Immunol. 198, 4156; DOI: https://doi.org/10.4049/jimmunol.1601921) and Boyer et al. (J. Immunol. 198, 4148; DOI: https://doi.org/10.4049/jimmunol.1601924).
This work was supported by EuroClonality.
- Received December 15, 2016.
- Accepted January 9, 2017.
- Copyright © 2017 by The American Association of Immunologists, Inc.