EuroFlow Quality Assessment program for EuroFlow protocols for the diagnosis of hematological malignancies

The EuroFlow Quality Assessment Program for EuroFlow Protocols used to diagnose hematological malignancies (the LST-QA program), to detect a bone marrow minimal residual disease in multiple myeloma (MM MRD-QA program) and to screen for primary immunodeficiencies (PIDOT-QA program).

Introduction

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, 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, Next Generation Flow cytometry for MM MRD approaches, and/or the EuroFlow Primary Immunodeficiency Orientation Tube). 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 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 EuroFlow Primary Immunodeficiency Orientation Tube (PIDOT)-QA program

The EuroFlow Primary Immunodeficiency Orientation Tube (PIDOT)-QA program is designed to provide an external control for multiparametric flow cytometry screening for primary immunodeficiencies. This program is divided in two parts:

  1. A ‘wet lab’ part, which is designed similarly to LST-QA program. Peripheral blood (PB) samples of three healthy donors are taken locally in each participating laboratory. The samples are stained with the PIDOT 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 PIDOT 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. 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.
  2. A ‘dry’ part, in which participants download fcs files prepared by the staff from peripheral blood of patients suffering from immunodeficiency. Participants analyse these files and report their conclusions. Their performance is then compared to the reference results and to other (anonymized) participants. The results are summarized in the PIDOT-QA Certificate.

The EuroFlow LST-QA, MM-MRD-QA and PIDOT-QA programs are organized in collaboration with Dr. T. Kalina (QA leader, Charles University, Prague, CZ), Dr. Juan Flores-Montero (University of Salamanca, Salamanca, ES) and Prof. Dr. Carolien Bonroy (University of Ghent, Ghent, BE) members of the EuroFlow Consortium.

The structure

The EuroFlow QA programs are organized twice a year. For reasons of convenience and efficiency, participants sign up to the LST-QA, MM-MRD-QA and PIDOT-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, a EuroFlow QA Educational meeting is organized. Participation of two lab members is free of charge for QA program participants.

EuroFlow QA subscription options:

  1. Participation in one LST-QA round: €200
  2. Participation for the whole year for LST-QA round (2 rounds): €350
  3. Participation in one MM-MRD QA round: €200
  4. Participation for the whole year for MM-MRD-QA round (2 rounds): €350
  5. Participation for one PIDOT-QA round: €200
  6. Participation for the whole year for PIDOT-QA round (2 rounds): €350

If you would like to receive multiple reports, e.g. to test several instruments, there is a 50% discount for additional certificates.

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

The EuroFlow QA schedule for the year 2022:

EuroFlow QA 2022-I

Registration for all programs: January 10 - February 28 (registration for both rounds will be possible during this period)
LST-QA and MM-MRD-QA program:
Results reporting through the website: March 1 - March 30
Certificates issued: April 29
PIDOT-QA program:
Results reporting through the website: April 1 - April 29
Certificates issued: May 31

EuroFlow QA 2022-II

Registration for all programs: June 1 - August 26
LST-QA and MM-MRD-QA program:
Results reporting through the website: August 29 - September 30
Certificates issued: October 31
PIDOT-QA program:
Results reporting through the website: October 3 - October 31
Certificates issued: November 30

EuroFlow QA educational meeting

The EuroFlow QA participants are invited to join the EuroFlow QA educational meeting where possible 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.

The EuroFlow QA educational meeting is scheduled for autumn 2022. Details will follow.

Literature

If you have questions concerning the QA program please contact euroflowqa@euroflow.org