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Chronic lymphocytic leukemia

Impact of using leader primers for IGHV mutational status assessment in chronic lymphocytic leukemia

Leukemia volume 34, pages 2257–2259 (2020)Cite this article

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For chronic lymphocytic leukemia (CLL) patients, the mutational status of the rearranged immunoglobulin heavy chain variable (IGHV) gene has a strong prognostic value [1, 2] and might impact the choice of first line treatment in the future [3]. Given the importance of this biomarker, the ERIC consortium (European Research initiative on CLL) has elaborated technical recommendations which aim to increase the performances of the analysis [4].

In the last version of the ERIC recommendations [4], the consortium has recommended to change the design of the polymerase chain reaction (PCR) primers. Most of the laboratories use FR1 primers based on the BIOMED-2 design [5], which are highly efficient to amplify the clonal rearrangement, but generate smaller amplicons than leader primers. By contrast, leader primers are reputed to be less efficient [6], but generate large amplicons which enables a more accurate determination of the mutational status. Whereas both set of primers were considered as acceptable in the 2007 version of the ERIC recommendation [6], the use of FR1 primers is presented as “non acceptable” in the 2017 update, because it could lead to underestimation of the mutational load [4]. Whereas primer leaders generate longer amplicons and accordingly allow the determination of the mutational status based on a longer sequence analysis, they fail to amplify the rearrangement in a fraction of the cases, which might limit their interest [6]. As far as we know, the impact of the use of both set of primers has never been quantified in a published series. In order to fill this knowledge gap, we have conducted a comparative study of both sets of primers in a retrospective cohort of patients from the Lyon university hospital hematology laboratory.

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Fig. 1: Proportion of patients with a percentage identity compared to germ-line in the gray zone 97–99%.
Fig. 2: Comparison of the results using FR1 and leader primers for determination of IGHV mutation status.

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Authors and Affiliations

  1. Hospices Civils de Lyon, Service d’hématologie biologique, Centre Hospitalier Lyon Sud, Pierre-Bénite, France

    Sarah Huet, Anne Bouvard, Isabelle Mosnier, Kaddour Chabane, Sandrine Hayette & Pierre Sujobert

  2. Centre de Recherche en Cancérologie de Lyon, UMR INSERM 1052 CNRS 5286, Equipe «Clinical and Experimental Models of Lymphomagenesis», Lyon, France

    Sarah Huet, Gilles Salles, Sandrine Hayette & Pierre Sujobert

  3. Université Claude Bernard Lyon I, Lyon, France

    Sarah Huet, Gilles Salles & Pierre Sujobert

  4. Hospices Civils de Lyon, Service d’hématologie clinique, Centre Hospitalier Lyon Sud, Pierre-Bénite, France

    Emmanuelle Ferrant & Gilles Salles

  5. Centre Léon Bérard, Service d’hématologie, Lyon, France

    Anne Sophie Michallet

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  1. Sarah Huet
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Correspondence to Sarah Huet.

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Huet, S., Bouvard, A., Ferrant, E. et al. Impact of using leader primers for IGHV mutational status assessment in chronic lymphocytic leukemia. Leukemia 34, 2257–2259 (2020). https://doi.org/10.1038/s41375-020-0716-1

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