Introduction

The diagnosis of malignant lymphoma is recognized as a difficult area in histopathology. Modern hematopathology includes molecular tests such as clonality assays, since in principle B-cell and T-cell lymphomas are clonal diseases. Between 5 and 15% of hematopathology cases may benefit from clonality testing due to uncertainties based on histological and immunophenotypical evaluation. The development of new primer sets for clonality testing by the BIOMED-2 consortium [1] has resulted in standardization and significantly improved detection of clonality of malignant B-cell lymphomas/leukemias with the Ig-primer sets [2] and of malignant T-cell lymphomas/leukemias with the TCR primer sets [3]. Because of their high detection rates (99% for both B-cell and T-cell malignancies) and the complementarity of the primer sets, these assays are now widely used for detection of clonality in lymphoproliferative disorders.

The original BIOMED-2 study group has continued collaboration and is now known as the EuroClonality consortium. EuroClonality focuses on educational activities for Ig/TCR clonality testing, on quality control and development of guidelines for performance and evaluation of diagnostic clonality testing. The EuroClonality members meet twice a year to discuss technical and interpretation problems on clinical cases, to study specific lymphoma subtypes, to develop new applications and methodologies and to promote further developments in the field of hemato-oncology including a common nomenclature for the description and reporting of results derived from clonality testing.

From questions by others during the last few years, it is clear that clonality testing, especially the evaluation of results for patient diagnosis, can be difficult. After design and evaluation of the BIOMED-2/EuroClonality assays, we have therefore taken the responsibility to further educate the users of the clonality tests by organizing workshops, by reporting educational and practical examples of clonality testing [4–6] and by offering clonality support via the EuroClonality website. The public part is accessible to the wider clonality testing community and can be used to obtain information, education or support about clonality testing in routine diagnostic practice. In addition, the website functions as a communication medium for the EuroClonality group itself (the non-public part that is password protected).

EuroClonality public website: information, workshop and FAQs

On the website, general information about the EuroClonality consortium and the main goals of the group are documented. In addition, there is information on the workshop: “Clonality Assessment in Pathology”, which is organized for laboratories that have introduced clonality testing using the BIOMED-2/EuroClonality primer sets. Currently many laboratories throughout the world are still introducing the technology, resulting in questions about the technology and interpretation of the results in routine practice. In addition, these laboratories wish to further improve the quality and reliability of their clonality assessment. For these laboratories, the EuroClonality consortium organizes annual workshops. Since the vision of the EuroClonality consortium is that clonality testing can only be performed in a reliable way when a close interaction exists between the (clinical) molecular biologist and the hematopathologist, these workshops are organized for such teams of specialists. Participants can download a registration form and send it to the workshop secretariat (workshop@euroclonality.org). Besides educational sessions, there are sessions in which individual cases are discussed. The participants can bring their own cases, which are difficult from the perspective of pathology and/or from the molecular results (GeneScan or gel based heteroduplex analysis of clonality results). All cases are discussed within the group in 20 headed microscope sessions, and knowledge and opinions are shared (Fig. 1). These workshops have proven to be very fruitful learning experiences for all participants. In the future we will consider organization of similar workshops for hematologists and immunologists.

Fig. 1
figure 1

Photographs taken during an educational session (a, b) and during the case presentations and discussions (c, d)

Full size image

Frequently asked questions (FAQs) can also be found on the website. There is already a list of questions from “detection rate of clonality tests” to “software used in clonality analysis” which have been answered by an expert team. Those who perform clonality analysis can use this information as an educational resource. The FAQs as they are currently available via the website are documented in Table 1.

Table 1 Frequently asked questions in clonality testing currently available via the website
Full size table

EuroClonality public website: online support service

EuroClonality offers an online support service for questions on cases that are not answered by the frequently asked questions section. With this service, scientists that perform clonality testing can submit a problematic clonality case by filling in an online support form with details of the case and results obtained. A review board of experts is available to discuss technical and interpretation problems on clonality assessment and will reply to the queries within 10 working days.

Logistics for the online support service

The submission of difficult cases occurs online via the EuroClonality web page. For this purpose, a short description of the differential diagnostic question and a compilation of the previous results can be uploaded together with representative images. After completion of the upload process, the expert panel will be notified that there is a new case submitted for review. Each case receives a unique ID number, to distinguish the different cases.

The expert on duty analyzes the support question and reviews the submitted data. Based on his or her opinion, an initial reply proposal is made to the other experts. This proposal is discussed within the expert team by e-mail communication. The members of the expert team respond as fast as possible with comments and additions. The expert on duty incorporates all of the expert opinions and makes a final answer which is sent to the support coordinator. The support coordinator sends an e-mail with the final answer to the case submitter on behalf of the expert team. This final answer is also published on the password-protected expert part of the website and can be used as a basis for other online support requests or for educational purposes within the EuroClonality group.

Most questions are related to interpretation problems involving IGH and IGL rearrangement results followed by questions regarding TCRG and TCRB PCR. A very typical question concerns the importance of dominant PCR products in isolated targets for the final interpretation of a given case. There is a significant degree of uncertainty whether such solitary PCR products are sufficient to conclude that the sample is monoclonal. A second important field of topics is related to more general technical questions such as suitable positive and negative controls, DNA amounts, DNA quality and non-specific bands. According to the experience of the expert panels, recommendations include additional analysis, because the use of insufficient controls, the application of a limited primer set and absence of duplicate results are the major obstacles observed. Most request submissions are derived from European countries, but some are from the USA and Canada.

In addition to the service for pathology laboratories, EuroClonality has now also started a support service for cases that are submitted from an immunological/hematological perspective. To this end experts with a background in immunology and/or hematology are available to answer questions and provide advice on further action. Scientists can upload their cases in the same way as for pathologically problematic cases and can provide details on PCR assay and flow-cytometry results. As this service has only recently been started, it has not been very actively consulted so far. However, it is expected that this will change in the near future.

EuroClonality non-public part for members and expert teams

There is also a private part of the website, which is password protected. This non-public part is available for EuroClonality members and is used for communication purposes. For example, documents and files are available with practical information from the meetings, such as minutes, guidelines and concepts or preview publications. Most of the non-public part is used for the web-based Quality Control rounds (QCs). These QCs are currently organized for members only, with the aim of designing new guidelines and a universal scoring system to improve clonality test reporting. In the near future, the possibility to widen the activities of the group, especially the QCs and to make it accessible to a broader public, will be considered.

Finally, there is a private part of the website for the review board experts only. Experts can download support cases together with the clonality result files. A graphical overview of the design of the website is presented in Fig. 2. The website is hosted by one.com and written in html and php.

Fig. 2
figure 2

Sitemap of the EuroClonality website

Full size image

Summary

The EuroClonality website offers several tools, such as the EuroClonality workshops, the Frequently Asked Questions (FAQ) and the online support service for questions on clonality assessment in lymphoma diagnostics. The website is open to every pathologist, clinical molecular biologist, hematologist or immunologist interested in the field of lymphoproliferations. By offering educational tools and support, EuroClonality continues to contribute to the improvement of molecular (clonality) diagnosis of malignant lymphomas and thereby to the better management of our patients.