Establishment of immunoglobulin heavy (IGH) chain clonality testing by next-generation sequencing for routine characterization of B-cell and plasma cell neoplasms Journal Article


Authors: Arcila, M. E.; Yu, W.; Syed, M.; Kim, H.; Maciag, L.; Yao, J.; Ho, C.; Petrova, K.; Moung, C.; Salazar, P.; Rijo, I.; Baldi, T.; Zehir, A.; Landgren, O.; Park, J.; Roshal, M.; Dogan, A.; Nafa, K.
Article Title: Establishment of immunoglobulin heavy (IGH) chain clonality testing by next-generation sequencing for routine characterization of B-cell and plasma cell neoplasms
Abstract: Immunoglobulin heavy chain (IGH) clonality testing by next-generation sequencing (NGS) offers unique advantages over current low-throughput methods in the assessment of B-cell lineage neoplasms. Clinical use remains limited because assays are not standardized and validation/implementation guidelines are not yet developed. Herein, we describe our clinical validation and implementation of NGS IGH clonality testing and summarize our experience based on extensive routine use. NGS-based clonality testing targeting IGH FR1, FR2, FR3, and the conserved leader sequence upstream of FR1 was validated using commercially available kits. Data were analyzed by commercial and in-house–developed bioinformatics pipelines. Performance characteristics were evaluated directly comparing with capillary electrophoresis (CE) assays (BIOMED-2 primers). Assays were monitored after implementation (>1.5 years), concurrently testing by CE methods. A total of 1189 clinical samples were studied (94 validation, 1095 postimplementation). NGS showed superior performance compared with CE assays. For initial assessment, clonality detection rate was >97% for all malignancy types. Concordance with CE was 96%; discordances were related to higher sensitivity/resolution of NGS and improved detection in cases with high somatic hypermutation. Routine NGS clonality assessment is feasible and superior to existing assays, enabling accurate and specific index clone assessment and future tracking of all rearrangements in a patient sample. Successful implementation requires new standardization, validation, and implementation processes, which should be performed as a multicenter and multidisciplinary collaboration. © 2019 American Society for Investigative Pathology and the Association for Molecular Pathology
Keywords: controlled study; human tissue; unclassified drug; major clinical study; plasmacytoma; medical record review; retrospective study; b lymphocyte; somatic hypermutation; dna; immunoglobulin heavy chain; conserved sequence; clonal variation; capillary electrophoresis; bioinformatics; health care planning; next generation sequencing; human; article; immunoglobulin heavy chain fr1; immunoglobulin heavy chain fr2; immunoglobulin heavy chain fr3
Journal Title: Journal of Molecular Diagnostics
Volume: 21
Issue: 2
ISSN: 1525-1578
Publisher: Elsevier Science, Inc.  
Date Published: 2019-03-01
Start Page: 330
End Page: 342
Language: English
DOI: 10.1016/j.jmoldx.2018.10.008
PUBMED: 30590126
PROVIDER: scopus
DOI/URL:
Notes: Article -- Export Date: 1 April 2019 -- Source: Scopus
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MSK Authors
  1. Khedoudja Nafa
    184 Nafa
  2. Jinjuan Yao
    24 Yao
  3. Jae Hong Park
    147 Park
  4. Christine Gi-Yun Moung
    10 Moung
  5. Ahmet Zehir
    157 Zehir
  6. Maria Eugenia Arcila
    350 Arcila
  7. Paulo A Salazar
    19 Salazar
  8. Ivelise A Rijo
    11 Rijo
  9. Ahmet Dogan
    174 Dogan
  10. Mikhail Roshal
    68 Roshal
  11. Carl Ola Landgren
    198 Landgren
  12. Tessara   Baldi
    11 Baldi
  13. Mustafa H Syed
    11 Syed
  14. Caleb   Ho
    23 Ho
  15. Wayne   Yu
    6 Yu
  16. Hannah Jung Kim
    2 Kim
  17. Lidia Maciag
    3 Maciag