EuroFlow Quality Assessment Program
The EuroFlow Quality Assessment Program for EuroFlow Protocols used to diagnose hematological malignancies (the LST-QA program) and to detect a bone marrow minimal residual disease in multiple myeloma (MM MRD-QA program)
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Introduction
The flow cytometric immunophenotyping has become essential for an accurate diagnosis, classification, and disease monitoring in hemato-oncology. The EuroFlow Consortium has established a fully standardized "all-in-one" pipeline consisting of standardized instrument settings, reagent panels, sample preparation protocols, and software for data analysis and disease classification. The software tools enable a large-scale data analysis and a direct comparison of individual cases against reference data sets composed of specific disease categories. The need for comparability of antigen expression measured in different centers called for a quality assessment (QA) program to ensure that the high level of standardization is maintained in all participating centers. Therefore, the EuroFlow developed innovative and easy to implement external QA programs which aim at monitoring the flow cytometric process (cytometer setting, sample preparation, data acquisition and/or data analysis).
EuroFlow QA programs are proficiency tests specific for laboratories that use standardized EuroFlow protocols (the participants should be routinely using the EuroFlow Lymphoid Screening Tube and/ or Next Generation Flow cytometry for MM MRD approaches). They may be used to complement, but not to replace, other established proficiency tests.
The EuroFlow Lymphoid Screening Tube (LST)-QA program
The EuroFlow LST-QA program is designed to mimic the routine sample preparation and data acquisition, as used for local diagnostic samples. For each LST-QA round, peripheral blood (PB) samples of three healthy donors are taken locally in each participating laboratory. The samples are stained with the LST antibody panel and measured on the local flow cytometer, strictly following the EuroFlow sample preparation standard operating procedure (SOP) and the EuroFlow SOP for instrument setting and fluorescence compensation. The FCS files are analysed in the Infinicyt software (Cytognos SL, Salamanca, ES) using a QA profile downloaded from the QA webpage and a recommended gating strategy (described in detail in the test instructions). Alternative analytical software can also be used, in which case it will not be possible to provide the participants with the graphical representation of their performance in relation to others. The results are reported via the QA website. The QA staff then compares the median fluorescence intensity (MedFI) values with the reference data. Deviations from the reference values are quantified using a performance score metrics and the results are summarized in the LST-QA Certificate.
The EuroFlow Next Generation Flow Multiple Myeloma Minimal residual disease (MM MRD)-QA program
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 patient 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 EuroFlow LST-QA and MM-MRD-QA programs are organized in collaboration with Dr. T. Kalina (QA leader, Charles University, Prague, CZ) and Dr. Juan Flores-Montero (University of Salamanca, Salamanca, ES), members of the EuroFlow Consortium.
If you have questions concerning the QA programs, please contact euroflowqa@euroflow.org
The structure
The EuroFlow QA programs are organized twice a year. For reasons of convenience and efficiency, participants sign up to the LST-QA and MM-MRD-QA rounds separately.
We provide an opportunity to sign-up to the two yearly QA rounds of each program at the beginning of the year (which will result in a single invoice). A separate subscription for a single round is also possible.
Once a year, EuroFlow QA educational meeting is organized, participation of two lab memebers is free of charge for QA program participants.
EuroFlow QA programs subscription options:
a) |
Participation in one LST-QA round |
€200 |
b) |
Participation for the whole year (2 LST-QA rounds) |
€350 |
c) |
Participation in one MM-MRD-QA round |
€200 |
d) |
Participation for the whole year (2 NGF-MM-MRD-QA rounds) |
€350 |
Each addtional certificate in the LST-QA program |
€90 |
The EuroFlow QA programs schedule for the year 2021:
EuroFlow QA 2021-I
Registration: January 11 - March 14 (registration for both rounds will be possible during this period)
(the participation cost invoice distribution and payment)
Results’ reporting through the website: April 5 - May 2
Certificates issued: May 31 – June 6
EuroFlow QA 2021-II
Registration: April 5 - June 6
(the participation cost invoice distribution and payment)
Results’ reporting through the website: September 2 - September 28
Certificates issued: October 25 – October 31
EuroFlow LST-QA educational meeting
Autumn 2021
The EuroFlow QA participants are invited to join the EuroFlow QA educational meeting where the potential problems and pitfalls will be discussed. Participation in the meeting is free of charge for 2 representatives per participating laboratory, including coffee breaks and lunches.
Invoices will be issued by the EuroFlow foundation:
EuroFlow
Dept. of IHB, LUMC,
Building 1, E3-Q, Room L1-71G
Albinusdreef 2
2333 ZA Leiden
The Netherlands
VAT Number: NL857822986B01
Contact person: W.M. Bitter
Email: w.m.bitter@eslho.org
T: +31 71 526 6871
Name of bank: Rabobank
IBAN code: NL66 RABO 0347 3180 37, in the name of Stichting EuroFlow
BIC or SWIFT code: RABONL2U
Tomas Kalina, Juan Flores-Montero and Nada Brdickova, EuroFlow QA organizers
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, https://doi.org/10.1016/j.jim.2018.09.008