FLAER-negative granulocytes detection in periferal blood is simple and highly reproducible test for pnh screening

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Abstract

GPI-deficient cells' detection by multicolor flow cytometry using fluorescently labeled aerolysin (FLAER) currently is a standard approach for PNH diagnostics. Nevertheless more simple and cheap approach with high sensitivity is needed for routine PNH screening. The aim of present study was to investigate the applicability of simplified two-color (FLAER/CD15) flow cytometry for PNH clone screening. Search for FLAER-negative granulocytes was performed in 259 peripheral blood samples obtained from the patients with suspected PNH. Symplified analysis results were compared to the multicolor data. Both in test cohort (n = 216) and validation cohort (n = 43) FLAER-negative granulocytes detection results correlated well with conventional cytometric test findings. Simplified approach diagnostic sensitivity achieved 100% while rate of false positive results was very low (2.63% in test cohort and 0% in validation cohort). Thus two-color detection of FLAER-negative I granulocytes is effective test as a screening for presence of PNH clone. Nevertheless the positive result of screening investigation should be always confirmed by multicolor assessment of PNH clone size among erythrocytes, granulocytes and monocytes.

About the authors

Alexander M. Popov

Federal Research Center for Pediatric Hematology, Oncology and Immunology

Author for correspondence.
Email: uralcytometry@gmail.com
Russian Federation

Svetlana A. Kashpor

Federal Research Center for Pediatric Hematology, Oncology and Immunology

Email: cldimmynol.fgu@mail.ru
Russian Federation

Tatiana Yu. Verzhbitskaya

Pediatric Oncology/Hematology Center, Regional Children Hospital, Ekaterinburg

Email: uralverba@gmail.com
Russian Federation

Vyacheslav V. Borisov

Alexion Pharma

Email: borisovv@alxn.com
Russian Federation

Larisa G. Fechina

Pediatric Oncology/Hematology Center, Regional Children Hospital, Ekaterinburg

Email: childrens_oncology@mail.ru
Russian Federation

Svetlana A. Plyasunova

Federal Research Center for Pediatric Hematology, Oncology and Immunology

Email: svetlana.plyasunova@fccho-moscow.ru
Russian Federation

Alexey A. Maschan

Federal Research Center for Pediatric Hematology, Oncology and Immunology

Email: info@fnkc.ru@mail.ru
Russian Federation

Galina A. Novichkova

Federal Research Center for Pediatric Hematology, Oncology and Immunology

Email: info@fnkc.ru@mail.ru
Russian Federation

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Copyright (c) 2016 Popov A.M., Kashpor S.A., Verzhbitskaya T.Y., Borisov V.V., Fechina L.G., Plyasunova S.A., Maschan A.A., Novichkova G.A.

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This work is licensed under a Creative Commons Attribution 4.0 International License.