Next Generation Flow™ solution

for Primary Immunodeficiencies

Download EuroFlow PID Workflow Poster
Download the white paper


Primary Immunodeficiency Orientation Tube (PIDOT)

  • Orientates to the probable diagnosis of Primary Immunodeficiencies following the ESID criteria (T-, B- and NK cells) and to the genetic defect to be confirmed by molecular biology.
  • The standardized panel was used to create a database that describes the frequency and absolute count of all cell populations that can be identified in a peripheral blood sample.
  • Functional and maturation specific markers with clinical value are included in this combination to identify quickly and unequivocally all cell subsets.
  • High sensitivity close to 10-6.
BV421* BV510* FITC PE PerCP-Cyanine5.5 PE-Cyanine7 APC APC-C750™
CD27 CD45RA CD8+SmIgD CD16+CD56 CD4+SmIgM CD19+TCRgd CD3 CD45

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Characterization for Primary Immunodeficiencies

n contrast to immunoglobulin levels in serum, more dynamic changes are observed at a cellular level for total B-cell numbers and relative distribution of the B-cell compartment in peripheral blood.

  • Age-related reference values are available when using this IgH isotype B-cell tube together with CD27, IgM, CD21, CD24, CD19, SmIgD, CD5, CD38.
    This combination give accurate and robust information even in infant low-volume samples.
  • Response to vaccination can be predicted in elderly patients with reduced ability to produce antigen-specific antibodies.
  • The evaluation of plasma cell (PC) counts can be used as a surrogate marker for future PC production in bone marrow.

EuroFlow™ has designed two versions of this panel, adapted to 8-color and 12-color flow cytometers, which differ on the fluorochrome-marker combinations and the number of tubes used.

12-color panel

FITC PE PerCP-Cyanine5.5 APC
Tube 1 IgG2+IgG3 IgG1+IgG2 IgA1+IgA2 IgA1+IgG4

8-color panel

Tube 1 IgG2+IgG4 IgG1+IgG2
Tube 2 IgA1+IgG3 IgA1+IgA2

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Automated Analysis:

  • EuroFlow™ Database containing normal peripherical blood samples.
  • The Reference Database includes samples collected from different centers reflecting biological and technical variability.
  • Automated identification of all normal counterparts and detection of abnormal cells reduces analysis time and makes the process user-independent.
  • Complete immune profile information is relevant for prognosis and an indicator of patients with sustained disease (e.g. high normal plasma cell recovery and an immune profile with increased B-cell maturation are indicators of good prognosis).

Automatic Report:

  • Automatic comparison of the frequency of each population with reference ranges from the EuroFlow™ Database.
  • Clinically relevant comments and conclusions based on reference values and patient specific-results.
  • Improves communication between clinicians and flow cytometry laboratories.
  • Available in different languages.
  • Results can be linked to your Laboratory Information System (LIS).

Learn more about the EuroFlow™ Databases

A strong technical support team with the scientific-based knowledge and practical experience to implement Next Generation Flow™

Cytognos Technical Support team have the required experience, know-how and training resources to achieve successful implementation of the NGF methodology independently of the site (public and private hospitals, research facilities or other). The following points are addressed during technical support:

  • SOPs for cytometer set-up: Instrument standardization introduced in all labs.
  • SOPs and stable lyophilized kits for sample processing: Inter and intra laboratory reproducibility.
  • Data analysis: Reference databases with standardized reports bring a common language to the different sites.

Technical support is available through email, webinars or onsite visits. Cytognos provides a variety of solutions and products specifically aimed at the establishment of Next Generation Flow™ in your lab. Feel free to contact us to know more about them.


  1. van der Burg M, et al. The EuroFlow PID Orientation Tube for Flow Cytometric Diagnostic Screening of Primary Immunodeficiencies of the Lymphoid System. Front Immunol. 2019;10:246. Go to publication
  2. van Dongen JJM, et al. EuroFlow-Based Flowcytometric Diagnostic Screening and Classification of Primary Immunodeficiencies of the Lymphoid System. Front Immunol. 2019;10:1271. Go to publication
  3. Blanco E, et al. Defects in memory B-cell and plasma cell subsets expressing different immunoglobulin-subclasses in patients with CVID and immunoglobulin subclass deficiencies. J Allergy Clin Immunol. 2019 Feb 28. pii: S0091-6749(19)30280-5. Go to publication
  4. Criado I, et al. Residual normal B-cell profiles in monoclonal B-cell lymphocytosis versus chronic lymphocytic leukemia. Leukemia. 2018;32(12):2701–05. Go to publication
  5. Blanco E, et al. Selection and validation of antibody clones against IgG and IgA subclasses in switched memory B-cells and plasma cells. J Immunol Methods. 2017 Sep 28. pii: S0022-1759(17)30079-0. Go to publication