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Comparative Study
. 2021 Aug;479(2):365-376.
doi: 10.1007/s00428-021-03046-0. Epub 2021 Mar 8.

Evaluation of a worldwide EQA scheme for complex clonality analysis of clinical lymphoproliferative cases demonstrates a learning effect

Cleo Keppens  1 , Elke Boone  2 , Paula Gameiro  3 , Véronique Tack  1 , Elisabeth Moreau  2 , Elizabeth Hodges  4 , Paul Evans  5 , Monika Brüggemann  6 , Ian Carter  7 , Dido Lenze  8 , Maria Eugenia Sarasquete  9 , Markus Möbs  8 , Hongxiang Liu  10 , Elisabeth M C Dequeker  1 , Patricia J T A Groenen  11
Affiliations
Comparative Study

Evaluation of a worldwide EQA scheme for complex clonality analysis of clinical lymphoproliferative cases demonstrates a learning effect

Cleo Keppens et al. Virchows Arch. 2021 Aug.

Abstract

Clonality analysis of immunoglobulin (IG) or T-cell receptor (TR) gene rearrangements is routine practice to assist diagnosis of lymphoid malignancies. Participation in external quality assessment (EQA) aids laboratories in identifying systematic shortcomings. The aim of this study was to evaluate laboratories' improvement in IG/TR analysis and interpretation during five EQA rounds between 2014 and 2018. Each year, participants received a total of five cases for IG and five cases for TR testing. Paper-based cases were included for analysis of the final molecular conclusion that should be interpreted based on the integration of the individual PCR results. Wet cases were distributed for analysis of their routine protocol as well as evaluation of the final molecular conclusion. In total, 94.9% (506/533) of wet tests and 97.9% (829/847) of paper tests were correctly analyzed for IG, and 96.8% (507/524) wet tests and 93.2% (765/821) paper tests were correctly analyzed for TR. Analysis scores significantly improved when laboratories participated to more EQA rounds (p=0.001). Overall performance was significantly lower (p=0.008) for non-EuroClonality laboratories (95% for IG and 93% for TR) compared to EuroClonality laboratories (99% for IG and 97% for TR). The difference was not related to the EQA scheme year, anatomic origin of the sample, or final clinical diagnosis. This evaluation showed that repeated EQA participation helps to reduce performance differences between laboratories (EuroClonality versus non-EuroClonality) and between sample types (paper versus wet). The difficulties in interpreting oligoclonal cases highlighted the need for continued education by meetings and EQA schemes.

Keywords: Clonality analysis; External quality assessment; IG rearrangements; TCR rearrangements.

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Conflict of interest statement

EuroClonality is a scientific foundation that, together with the foundations EuroMRD and EuroFlow, is connected to the ESLHO foundation/EHA scientific working group. The objectives of the EuroClonality Foundation are aimed at innovation, standardization, quality control, and education in the field of diagnostic clonality analysis. The revenues of the previously obtained patent (PCT/NL2003/000690), which is collectively owned by the EuroClonality Foundation and licensed to Invivoscribe, are exclusively used for EuroClonality activities, such as for covering costs of the Working Group meetings, collective WorkPackages (including the WorkPackage: External Quality Control, EQA), and the EuroClonality Educational Workshops.

- Cleo Keppens has nothing to declare.

- Elke Boone is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2014–now).

- Paula Gameiro is a member of the Board of the EuroClonality Scientific Foundation, and chair of the WorkPackage EQA (assessor 2014–now).

- Véronique Tack has nothing to declare.

- Elisabeth Moreau is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2015–2018).

- Elizabeth Hodges is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2014–now).

- Paul Evans is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2017–now), and has received honoraria and lecture fees from Novartis, Diceutics, and Astellas within the past 36 months, outside the scope of this research.

- Monika Brüggemann is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2014–2016), performed contract research for Affimed, Amgen, Regeneron, advisory board of Amgen, Incyte, and is a speaker for the bureau of Janssen, Pfizer, Roche.

- Ian Carter is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2017–2018).

- Dido Lenze is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2015–2017).

- Maria Eugenia Sarasquete is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (2019–now).

- Markus Möbs is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (2019–now).

- Hongxiang Liu is a member of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2017–now).

- Elisabeth MC Dequeker has nothing to declare.

- Patricia JTA Groenen is chair of the EuroClonality Scientific Foundation and member of the WorkPackage EQA (assessor 2014–now).

Figures

Fig. 1
Fig. 1
Overview of the EuroClonality EQA scheme process. EQA, external quality assessment; IG, immunoglobulin gene; TR, T-cell receptor gene
Fig. 2
Fig. 2
Improvement upon repeated EQA participation for the different targets, sample, and participant types. #, number; IG, immunoglobulin gene; SEM, standard error of the mean; TR, T-cell receptor gene. Bar numbers represent the number of unique laboratories for which the average analysis score is calculated. No standard errors are shown for bars with only one participant
See this image and copyright information in PMC

References

    1. Van Dongen JJM, Wolvers-Tettero IL. Analysis of immunoglobulin and T cell receptor genes, part II. Clin Chim Acta. 1991;198:93–174. doi: 10.1016/0009-8981(91)90247-A. - DOI - PubMed
    1. Van Krieken JHJM, Langerak AW, Macintyre EA, et al. Improved reliability of lymphoma diagnostics via PCR-based clonality testing. Report of the BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia. 2007;21:201–206. doi: 10.1038/sj.leu.2404467. - DOI - PubMed
    1. Van Dongen JJ, Langerak AW, Bruggemann M, et al. Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia. 2003;17:2257–2317. doi: 10.1038/sj.leu.2403202. - DOI - PubMed
    1. Brüggemann M, White H, Gaulard P, Garcia-Sanz R, Gameiro P, Oeschger S, Jasani B, Ott M, Delsol G, Orfao A, Tiemann M, Herbst H, Langerak AW, Spaargaren M, Moreau E, Groenen PJTA, Sambade C, Foroni L, Carter GI, Hummel M, Bastard C, Davi F, Delfau-Larue MH, Kneba M, van Dongen JJM, Beldjord K, Molina TJ. Powerful strategy for PCR-based clonality assessment in T-cell malignancies. Report of the BIOMED-2 Concerted Action BHM4-CT98-3936. Leukemia. 2007;21:215–221. doi: 10.1038/sj.leu.2404481. - DOI - PubMed
    1. Evans PAS, Pott C, Groenen PJTA, Salles G, Davi F, Berger F, Garcia JF, van Krieken JHJM, Pals S, Kluin P, Schuuring E, Spaargaren M, Boone E, González D, Martinez B, Villuendas R, Gameiro P, Diss TC, Mills K, Morgan GJ, Carter GI, Milner BJ, Pearson D, Hummel M, Jung W, Ott M, Canioni D, Beldjord K, Bastard C, Delfau-Larue MH, van Dongen JJM, Molina TJ, Cabeçadas J. Significantly improved PCR-based clonality testing in B-cell malignancies by use of multiple immunoglobulin gene targets. Report of the BIOMED-2 Concerted Action BHM4- CT98-3936. Leukemia. 2007;21:207–214. doi: 10.1038/sj.leu.2404479. - DOI - PubMed

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