Mepacrine Flow Cytometry Assay for the Diagnosis of Platelet δ-granule Defects: Literature Review on Methods—Towards a Shared Detailed Protocol

Mehdi Khourssaji; Marion Bareille; Lorenzo Alberio; Delphine Borgel; Marc Fouassier; Marie-Christine Béné; Thomas Lecompte; François Mullier

doi:10.1055/a-2413-2870

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2025; 125(07): 618-633
DOI: 10.1055/a-2413-2870

Review Article

Mepacrine Flow Cytometry Assay for the Diagnosis of Platelet δ-granule Defects: Literature Review on Methods—Towards a Shared Detailed Protocol

Mehdi Khourssaji

¹Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium

²Institut de Recherche Expérimentale et Clinique (IREC) – Pôle Mont, Université catholique de Louvain, Yvoir, Belgium

,

Marion Bareille

¹Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium

²Institut de Recherche Expérimentale et Clinique (IREC) – Pôle Mont, Université catholique de Louvain, Yvoir, Belgium

,

Lorenzo Alberio

³Division of Haematology and Central Haematology Laboratory, Lausanne University Hospital (CHUV) and University of Lausanne (UNIL), Lausanne, Switzerland

,

Delphine Borgel

⁴Service d'Hématologie Biologique, Hôpital Necker AP-HP, Paris, France

,

Marc Fouassier

⁵Centre de Ressources et de Compétences - Maladies Hémorragiques Constitutionnelles, CHU de Nantes, Nantes, France

,

Marie-Christine Béné

⁶CRC2INA, Université de Nantes, Nantes, France

,

Thomas Lecompte

¹Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium

⁷Hematology Department and Grand East Competence Center on Inherited Platelet Disorders, CHU Nancy, Nancy, France

,

François Mullier

¹Department of Laboratory Medicine, Hematology Laboratory, CHU UCL Namur, Yvoir, Belgium

²Institut de Recherche Expérimentale et Clinique (IREC) – Pôle Mont, Université catholique de Louvain, Yvoir, Belgium

⁸Université de Namur, Department of Pharmacy, Namur Thrombosis and Hemostasis Center (NTHC), Namur Research Institute for Life Sciences (NARILIS), Namur, Belgium

› Author Affiliations

Abstract

Accurate assessment of platelet secretion is essential for the diagnosis of inherited or acquired platelet function disorders and more specifically in identifying δ-storage pool disease. Mepacrine, a fluorescent dye, specifically accumulates in platelet δ-granules. The mepacrine flow cytometry (mepacrine FCM) assay has been used for more than half a century in the clinical laboratory as a diagnostic tool for platelet δ-granule disorders. The assay requires a small volume of blood, can be performed in thrombocytopenic patients, provides rapid assessment of δ-granule content and secretion, and, thus, enables differentiation between storage and release defects. There is however a broad heterogeneity in methods, reagents, and equipment used. Lack of standardization and limited data on analytical and clinical performances have led the 2022 ISTH SSC (International Society on Thrombosis and Haemostasis Scientific and Standardization Committee) Subcommittee on Platelet Physiology expert consensus to rate this assay as simple but of uncertain value. Yet, the data used by experts to formulate the recommendations were not discussed and even not mentioned. Guidance for laboratory studies of platelet secretion assay would be very helpful for clinical laboratories and health authorities especially considering the implications of the new In Vitro Diagnostic Regulation in Europe. The purpose of the present work was to review the reported methodologies for the mepacrine FCM assay and to offer an example of detailed protocol. This would help standardization and pave the way for more rigorous comparative studies.

Keywords

mepacrine - flow cytometry - storage pool disease - platelet - inherited or acquired diseases - delta granule

Full Text

References

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