EuroFlow External Quality Assessment Program

The EuroFlow Consortium has established fully standardized methods for flow cytometry consisting of standardized instrument settings, reagent panels, sample preparation protocols, and software for data analysis and disease classification. The software tools enable large-scale data analysis and a direct comparison of individual cases against reference datasets composed of specific disease categories. The need for comparability of antigen expression measured in different centers calls for quality assessment to ensure that the high level of standardization is maintained in all participating centers. This also improves the reliability of the EuroFlow reference data. Therefore, EuroFlow developed innovative and easy to implement external quality assessment (EQA) schemes that aim at monitoring your flow cytometric process (cytometer settings, sample preparation, data acquisition, and/or data analysis an interpretation) and supporting your lab in implementing the EuroFlow protocols to the highest standard. The EuroFlow EQA program is meant to complement, but not to replace other established proficiency tests.

EQA for EuroFlow Protocols

The EQA schemes of EuroFlow are proficiency tests for laboratories that follow the EuroFlow protocols. The participants should routinely use the EuroFlow Lymphoid Screening Tube (LST), Multiple Myeloma (MM) MRD tube, B Cell Precursor-Acute Lymphocytic Leukemia (BCP-ALL) MRD tube, and/or the Primary Immunodeficiency Orientation Tube (PIDOT). The EuroFlow QA program uses local healthy donor samples and therefore no sample shipment is necessary.

EuroFlow offers four different EQA schemes:

The EuroFlow Lymphoid Screening Tube (LST)-QA program

This scheme is designed to mimic the routine sample preparation and data acquisition, as used in your lab. For each LST-QA round, peripheral blood (PB) samples obtained locally from three healthy donors are processed according to the EuroFlow method. The EQA provider then compare your results with the reference data. Deviations from the reference values are quantified using a performance score metrics and the results are summarized in a certificate.

The Multiple Myeloma (MM) Minimal Residual Disease (MRD)-QA scheme

The EuroFlow Next Generation Flow (NGF) Multiple Myeloma (MM) Minimal Residual Disease (MRD)-QA program is designed to provide an external control for multiparametric flow cytometry detection of MRD in MM patients after therapy. For each MM MRD-QA round, the QA staff prepares four FCS files to be downloaded by the participants. These FCS files represent bone marrow samples from MM patients after treatment, processed following the NGF methodology. Participants analyze each file and report their conclusions via an online form. Their performance is then compared to the reference results and to other (anonymized) participants. The results are summarized in the MM MRD-QA certificate.

The Primary Immunodeficiency Orientation Tube (PIDOT)-QA scheme

This scheme is designed to provide an external control for multiparametric flow cytometry screening for primary immunodeficiencies. It is divided into two parts, a ‘wet lab´ part, which is designed similarly to the LST-QA program and a ‘dry’ part, in which participants download FCS files prepared by the EQA provider using peripheral blood of patients suffering from immunodeficiency. Participants analyze these files and report their conclusions. Their performance is then compared to the reference results and to other (anonymized) participants.

The B Cell Precursor-ALL Minimal Residual Disease (BCP-ALL MRD)-QA scheme

This scheme combines a ‘wet lab´ part, which is similar to the LST-QA program, and a ‘dry’ part, which uses files from actual BCP-ALL patients, prepared by the EQA provider. Participants download these files, merge them with the control files obtained in the ‘wet lab´ part and analyze them together. This requires software that enables merging of the files (e.g. Infinicyt). The performance is then compared to the reference results and to other (anonymized) participants.

Feedback

After each EQA round you will be provided a personalized certificate in which you can clearly see how your results compare to the reference values, but also how you perform in relation to the other participating laboratories. The EQA provider not only provides you with clear data, but also with tips. The EuroFlow EQA schemes are concluded by a EuroFlow EQA Educational meeting. This meeting is organized once a year and participation is free of charge for two representatives per participating laboratory, including coffee breaks and lunches. During the meeting, possible problems and pitfalls will be discussed and you will have the opportunity to ask your questions.

Structure

The EuroFlow EQA schemes are organized in collaboration with:

• Prof. Dr. Tomas Kalina (QA leader for LST-QA; Charles University, Prague, CZ)
• Dr. Juan Flores-Montero (MM MRD-QA; University of Salamanca, Salamanca, ES).
• Prof. Dr. Carolien Bonroy (PIDOT-QA; University of Ghent, Ghent, BE)
• Dr. Michaela Reiterova (BCP-ALL MRD-QA; Charles University, Prague, CZ),

and other members of the EuroFlow Consortium.

The EuroFlow EQA schemes are organized twice a year. Participants may choose the EQA schemes according to their needs and interests.

EuroFlow QA subscription options:

Participation in one round of any QA scheme: € 250,- 

Participation for the whole year for one QA Scheme: € 425,-

For each additional certificate in the LST-QA program (e.g. for additional instruments) there will be an extra charge of €90. Other programs do not have the option of multiple reports.

You can register or log in on the EuroFlow QA web page.

The EuroFlow QA programs schedule for the year 2024:

Literature

• Kalina, T. et al. Quality Assessment Program for EuroFlow Protocols: Summary Results of Four-Year (2010-2013) Quality Assurance Rounds. Cytometry A. 87(2):145-56 (2015).
• Flores-Montero J. et al. Next Generation Flow for highly sensitive and standardized detection of minimal residual disease in multiple myeloma. Leukemia. 31(10):2094-2103 (2017).
• Kalina, T. et al. EuroFlow standardization of flow cytometer instrument settings and immunophenotyping protocols. Leukemia 26, 1986-2010 (2012).
• van Dongen, J. J. M. et al. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia 26, 1908-75 (2012).
• Kalina, T. et al. Frequent issues and lessons learned from EuroFlow QA. Journal of Immunological Methods 475, 112520 (2019)