Ann Lab Med 2019; 39(2): 150-157  https://doi.org/10.3343/alm.2019.39.2.150
Comparison of High Sensitivity and Conventional Flow Cytometry for Diagnosing Overt Paroxysmal Nocturnal Hemoglobinuria and Detecting Minor Paroxysmal Nocturnal Hemoglobinuria Clones
Sang Hyuk Park, M.D., Ph.D.1, Joseph Jeong, M.D., Ph.D.1, Seon-Ho Lee, M.D., Ph.D.1, Dong-Won Yoo, M.D.2, Yunsuk Choi, M.D., Ph.D.3, Jae-Cheol Jo, M.D., Ph.D.3, and Ji-Hun Lim, M.D., Ph.D.1
1Department of Laboratory Medicine, University of Ulsan College of Medicine, Ulsan University Hospital, Ulsan, Korea; 2Department of Laboratory Medicine, Pusan National University Yangsan Hospital, Yangsan, Korea; 3Department of Hematology and Cellular Therapy, University of Ulsan College of Medicine, Ulsan University Hospital, Ulsan, Korea
Corresponding author: Ji-Hun Lim, M.D. https://orcid.org/0000-0002-8205-9975
Department of Laboratory Medicine, University of Ulsan College of Medicine, Ulsan University Hospital, 877 Bangeojinsunhwando-ro, Dong-gu, Ulsan 44033, Korea
Tel: +82-52-250-7274
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E-mail: uucm77@gmail.com
Received: January 9, 2018; Revised: March 15, 2018; Accepted: September 20, 2018; Published online: March 1, 2019.
© Korean Society for Laboratory Medicine. All rights reserved.

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background: High sensitivity flow cytometry (HS-FCM) was recently developed for diagnosing paroxysmal nocturnal hemoglobinuria (PNH). We compared its performance with conventional flow cytometry (C-FCM) for diagnosing overt PNH and detecting minor (0.1–1%) PNH clones in aplastic anemia (AA)/low-grade myelodysplastic syndrome (MDS) patients.
Methods: C-FCM and HS-FCM were performed simultaneously on 41 samples from healthy controls and 23 peripheral blood samples from 15 AA/low-grade MDS and eight PNH patients, using a Navios flow cytometer (Beckman Coulter, Miami, FL, USA). Results were compared.
Results: No healthy control samples had PNH clone size >0.01%. For granulocytes, C-FCM detected a smaller PNH clone size than HS-FCM (mean difference: 0.7–1.7%). In AA/low-grade MDS patients, three samples showed >1% PNH clones with C-FCM but not with HS-FCM. Seven samples showed minor PNH clones by C-FCM, but HS-FCM showed negative results for all these samples. In PNH patients, C-FCM detected a smaller PNH clone size than HS-FCM (mean difference: 1.9–5.0%). For red blood cells, C-FCM detected a greater PNH clone size than HS-FCM (mean difference: 1.5%). In AA/low-grade MDS patients, C-FCM showed >1% PNH clones in six samples, but HS-FCM showed >1% PNH clones in none of the samples. C-FCM detected minor PNH clones in nine samples, but six of them were negative by HS-FCM. In PNH patients, C-FCM detected a greater PNH clone size than HS-FCM (mean difference: 2.5%).
Conclusions: HS-FCM can sensitively detect minor PNH clones and reduce false-positive C-FCM minor PNH clone cases in AA/low-grade MDS patients.
Keywords: Flow cytometry, High sensitivity, Paroxysmal nocturnal hemoglobinuria, Performance, Clone



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