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Related Experiment Video

Updated: Mar 6, 2026

Basophil Activation Test for Allergy Diagnosis
07:22

Basophil Activation Test for Allergy Diagnosis

Published on: May 31, 2021

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Basophil activation test: Implementation and standardization between systems and between instruments.

Anne-Emmanuelle Depince-Berger1, Khaled Sidi-Yahya1, Mohammed Jeraiby1

  • 1Immunology laboratory/University Hospital Saint-Etienne, FRE-CNRS 3312, 42055, Saint-Etienne Cedex 2, France.

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
|March 11, 2017
PubMed
Summary

Standardizing the basophil activation test (BAT) improves allergen hypersensitivity measurement. This study introduces a simplified, validated method for consistent, reliable results across different instruments and systems.

Keywords:
allergybasophil activation testdegranulationflow cytometrystandardization

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Area of Science:

  • Immunology
  • Flow Cytometry
  • Allergen Testing

Background:

  • The basophil activation test (BAT) is a valuable ex vivo method for assessing allergen hypersensitivity.
  • Current BAT protocols lack standardization, hindering inter-laboratory and inter-instrument reproducibility.

Purpose of the Study:

  • To implement and propose a standardized method for basophil activation and labeling using whole blood in EDTA.
  • To achieve inter-system and inter-instrument standardization for BAT.

Main Methods:

  • A simplified one-step stimulation and labeling method using BasoflowEx® and FlowCast® on EDTA whole blood.
  • Instrument setup standardization using Mean Fluorescence Intensity (MFI) targets with standard beads.
  • Validation according to ISO15189 standards, including precision and linearity assessments.
  • Co-labeling with B cells (CD20+) to assess relative basophil counts.

Main Results:

  • A simplified CD203c/CD63 (BasoflowEx) method for EDTA samples was established.
  • Optimized washing, concentration, and time parameters maximized basophil acquisition while minimizing noise and acquisition time.
  • The modified method demonstrated good precision (5.1% RCV) and linearity.
  • Standardization between different flow cytometry systems (Navios® and FACSCanto™) and instruments was achieved using MFI targets.
  • B cell co-labeling revealed potential underestimation of basophil responses in cases of strong hypersensitivity.

Conclusions:

  • The proposed method offers easy implementation for BAT, including EDTA use, one-step processing, and time-parameter analysis.
  • Standardization of instrument settings on MFI targets is crucial for reliable BAT results across various systems and instruments.
  • Standardized BAT can enhance the accuracy and reproducibility of hypersensitivity testing in sensitized patients.