External Quality Assessment Scheme for IG/TR Clonality Testing in Suspected Lymphoproliferations/Lymphoma

Information for Participants

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Introduction

In many hemato-oncology and pathology laboratories IG/TR clonality testing is part of the daily diagnostic work-up for samples with suspected lymphoproliferations or lymphoma. Standardization of PCR protocols for the detection of clonal proliferations in lymphoproliferative disease was established by the BIOMED-2 group (JJM van Dongen et al, Leukemia 2003:17:2257-2317, for literature and protocols see EuroClonality website,FAQsExit this website). These protocols are now well established in many labs, partly due to their commercial availability. These kits have made the technical performance of these protocols relatively uncomplicated, however the technical interpretation is less straightforward, since the reporting of these assays requires knowledge of T-cell receptor (TR) and immunoglobulin (IG) gene rearrangements, multiple rearrangements patterns and cross-lineage rearrangements.

To improve the standardization and reporting of clonality analysis, the Euroclonality group, which comprises more than 20 laboratories across Europe, with considerable technical experience of the BIOMED-2 primers and protocols, started a series of quality assessment (QA) schemes in 2008.
Two different kind of QA rounds were organized: paper trials and trials with DNA extracted from different hematological and histological cases, which participants analyzed according to their routine diagnostic PCR practice (Genescan and/or heteroduplex analysis) based on the suspected diagnosis. Paper trials focused on the interpretation of Genescan and/or heteroduplex results, whereas the DNA trials assessed both the correct generation of PCR-results as well as their interpretation. Participants were asked to submit results for individual loci tested, give an overall molecular interpretation and a final clinical interpretation where appropriate. These schemes resulted in the published guidelines for the analysis, interpretation and reporting of the Euroclonality/BIOMED-2 assays (Langerak, et al., Leukemia 2012;26:2159-2172, see FAQsExit this website EuroClonality website). To further standardize clonality reporting, language specific translations of these guidelines are available at the EuroClonality website. However, as these guidelines start from a immunobiological concept, it should be stressed that they can be applied to any clonality result whether or not generated with BIOMED-2 protocols.

Likewise for this scheme, both paper based and DNA based cases will be distributed annually. Participants need to test and/or interpret the EQA samples according to their laboratory’s routine practice, with the aim to improve and standardize diagnostic practices.

The practical organization of this European EQA program is done in collaboration with Dr Els Moreau and Dr Elke Boone, QA lead for Euroclonality and the Biomedical Quality Assurance Research Unit of the University of Leuven lead by Prof Dr Els Dequeker. The scheme organizers are members of this European working group and will be in close contact with the European QA program coordinator.

 

Set up of the scheme and the kind of samples distributed

In this EQA scheme, the tasks of preparing and sending the samples are assigned to laboratories within the Euroclonality network, who are in close contact with the Euroclonality QC lead, who with the Euroclonality board have the final responsibility for the EQA scheme.

Samples are presented as clinical cases and relevant clinical, flow cytometry and/or immunostaining data will be provided. As both hemato-oncology and pathology labs perform clonality analysis, sometimes for different clinical investigations, typical sample types (e.g. peripheral blood or FFPE tissue) will be included on a regular basis.

Five IG cases and five TR cases will be distributed together annually. Each trial round will be composed of 5 paper based cases and 5 DNA samples extracted from a variety of different lymphoproliferations. A rotation system will be applied so that after 2 years 5 paper based and 5 DNA samples for the IG scheme will be send around. A similar rotation system will be applied for the TR scheme.

For the 2016 IG and TR QA schemes participants will receive following cases:

  • 2016 IG scheme: 2 DNA samples and 3 paper based cases
  • 2016 TR scheme: 3 DNA samples and 2 paper based cases

The materials distributed are provided as specimens for the sole purpose of enabling external quality assessment at the recipient’s laboratory during the current distribution and must not be tested for any other target than that which is requested by the EQA scheme. They do not contribute in vitro medical diagnostic devices (IVDs), and no claim is made that they may be suitable for any other purpose or at any other point in time.

The scheme organizers prepare the samples and distribute them to the participants. Cases are pretested by minimal 3 laboratories of the EuroClonality group to determine the correct result. Co-ordination and evaluation of the results is done by the Euroclonality EQA scheme co-ordination center, as well as the Euroclonality QC lead.

 

Registration and result submission

Laboratories interested to participate in the EuroClonality EQA Scheme need to fill in this formExit this website.

Laboratories can register either for the IG trial, the TR trial or both. A fee of 400€ per gene target (IG or TR) or 800€ for both will be asked to the participants for the organization cost, preparation and sending of the samples and assessment of the results.

After registration each participant will receive a username and password which will allow them to log in to the EQA participants area where both sample information and links for result submission can be found.

The registrations are closed for the 2016 round. After closure of registration, participants will be notified by mail of their participation and will receive an invoice from the EuroClonality group.

Each year, all laboratories that have an account will receive an invitation via email to register online for the subsequent schemes. An identification number, the EQA ID number, is assigned to each participant automatically upon registration.

 

Evaluation

The coordination center in Leuven will coordinate the evaluation of the submitted results. Results of the EQA schemes will be announced after discussion in the EuroClonality working group. These results will be made available anonymously among the participants but each participating laboratory will receive individual feedback.

Scoring will initially be done on the final “Molecular Interpretation/Conclusion” of each case and not on the interpretation of the individual PCR-results, allowing laboratories who use non-BIOMED-2 protocols also to participate. However, in the final report results of BIOMED-2 PCR targets will be discussed in detail and feedback will be given.

The correct “Molecular Interpretation/Conclusion” was determined by minimal 3 EuroClonality laboratories during a pretesting phase. However, a case might be withdrawn from the EQA scoring in case high discordant results are obtained by the participants.

The IG/TR clonality EQA Scheme allows participants to react on the score and/or comments they received. All appeals should be sent via e-mail before the appealing deadline that is indicated in the general report. The appeals will be collected and will be discussed by the assessors. The laboratories will receive an individual answer and after this, the marks become final.

All participants receive a certificate of participation.
Laboratories will receive a certificate of good performance for either IG or TR analysis or both, after evaluation of 10 samples. After two years, at least 9 out of 10 cases should have been correctly assessed as determined by the “European Consensus Conference for External Quality Assessment in Molecular Pathology”Exit this website (van Krieken, et al., Virchows Arch 2013;462:27-37).
Laboratories that participated successfully, can be listed on the EuroClonality website if desired.

 

Timeline EQA 2016

  • May 17th, 2016 – June 17th, 2016: The registrations are closed for the 2016 round
  • End of August 2016: Distribution of the samples (you will receive an email when the samples are sent)
  • Deadline for submission of results: end of September 2016
  • November 2016: Results will be discussed with the EuroClonality group
  • December – January 2016: Report and individual results will be communicated to each participating laboratory

 

Successful participants EQA 2014

 

Successful participants EQA 2015

 

Confidentiality

The fact of participation, the raw data and the individual report are confidential information between the individual laboratory and the scheme organizer as well as the European QA coordinator.



Last changed: 2016-07-19