The EuroFlow PID Orientation Tube for Flow Cytometric Diagnostic Screening of Primary Immunodeficiencies of the Lymphoid System

Mirjam van der Burg, Tomas Kalina, Martin Perez-Andres, Marcela Vlkova, Eduardo Lopez-Granados, Elena Blanco, Carolien Bonroy, Ana E. Sousa, Anne Kathrin Kienzler, Marjolein Wentink, Ester Mejstríková, Vendula Šinkorova, Jan Stuchly, Menno C. van Zelm, Alberto Orfao, Jacques J.M. van Dongen

Research output: Contribution to journalArticleResearchpeer-review

58 Citations (Scopus)


In the rapidly evolving field of primary immunodeficiencies (PID), the EuroFlow consortium decided to develop a PID orientation and screening tube that facilitates fast, standardized, and validated immunophenotypic diagnosis of lymphoid PID, and allows full exchange of data between centers. Our aim was to develop a tool that would be universal for all lymphoid PIDs and offer high sensitivity to identify a lymphoid PID (without a need for specificity to diagnose particular PID) and to guide and prioritize further diagnostic modalities and clinical management. The tube composition has been defined in a stepwise manner through several cycles of design-testing-evaluation-redesign in a multicenter setting. Equally important appeared to be the standardized pre-analytical procedures (sample preparation and instrument setup), analytical procedures (immunostaining and data acquisition), the software analysis (a multidimensional view based on a reference database in Infinicyt software), and data interpretation. This standardized EuroFlow concept has been tested on 250 healthy controls and 99 PID patients with defined genetic defects. In addition, an application of new EuroFlow software tools with multidimensional pattern recognition was designed with inclusion of maturation pathways in multidimensional patterns (APS plots). The major advantage of the EuroFlow approach is that data can be fully exchanged between different laboratories in any country of the world, which is especially of interest for the PID field, with generally low numbers of cases per center.

Original languageEnglish
Article number246
Number of pages11
JournalFrontiers in Immunology
Publication statusPublished - 4 Mar 2019


  • automated gating strategy
  • EuroFlow
  • flow cytometric immunophenotyping
  • primary immunodeficiencies
  • standardization

Cite this