Performance Characteristics of a Non-Fluorescent Aerolysin-Based Paroxysmal Nocturnal Hemoglobinuria (PNH) Assay for Simultaneous Evaluation of PNH Neutrophils and PNH Monocytes by Flow Cytometry, Following Published PNH Guidelines
Language English Country United States Media print-electronic
Document type Journal Article
PubMed
27294344
DOI
10.1002/cyto.b.21389
Knihovny.cz E-resources
- Keywords
- CD157, PNH, flow cytometry, non-FLAER-based approach,
- MeSH
- ADP-ribosyl Cyclase immunology metabolism MeSH
- CD24 Antigen immunology metabolism MeSH
- Bacterial Toxins MeSH
- Biomarkers metabolism MeSH
- Antigens, CD immunology metabolism MeSH
- Pore Forming Cytotoxic Proteins MeSH
- GPI-Linked Proteins immunology metabolism MeSH
- Humans MeSH
- Lipopolysaccharide Receptors immunology metabolism MeSH
- Monocytes immunology metabolism MeSH
- Neutrophils immunology metabolism MeSH
- Hemoglobinuria, Paroxysmal immunology metabolism MeSH
- Flow Cytometry methods MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- ADP-ribosyl cyclase 2 MeSH Browser
- ADP-ribosyl Cyclase MeSH
- aerolysin MeSH Browser
- CD24 Antigen MeSH
- Bacterial Toxins MeSH
- Biomarkers MeSH
- Antigens, CD MeSH
- Pore Forming Cytotoxic Proteins MeSH
- GPI-Linked Proteins MeSH
- Lipopolysaccharide Receptors MeSH
BACKGROUND: CD157 has been recently reported as a useful glycosylphosphatidylinositol (GPI)-linked marker for the detection of paroxysmal nocturnal hemoglobinuria (PNH) clones in patients with suspected paroxysmal nocturnal hemoglobinuria by flow cytometry as it targets both neutrophils and monocytes. The aim of this study is to test the feasibility of a non-fluorescent aerolysin (FLAER) approach and propose an alternative for laboratories, where FLAER is not available. METHODS: We validated a non-FLAER-based single-tube, 6-color assay targeting the GPI-linked structures CD157, CD24, and CD14. We determined its performance characteristics on 20 PNH patient samples containing a variety of clone sizes and compared results with a previously validated FLAER-based approach. RESULTS: Coefficient of variation (CV) for intra-/interassay precision analyses ranged from 0.1%/0.2% to 3.02%/7.58% for neutrophils and from 0.10%/0.3% to 5.39%/6.36% for monocytes. Coefficient of determination (r2 ) for linear regression analysis of PNH clones from 20 patients ranging from 0.06% to 99.7% was 0.99 in all cases, Wilcoxon ranks test showed no statistically significant differences (P > 0.05), Bland-Altman analysis demonstrated performance agreement with mean bias ranging from 0.06 to 0.2. CONCLUSION: Our results confirm very good performance characteristics for both intra- and interassay precision analyses, favorable correlation, and agreement between the FLAER and non-FLAER-based approaches, using the CD157 GPI marker. Our experience suggests that a rapid and cost-effective simultaneous evaluation of PNH neutrophils and monocytes by flow cytometry without using FLAER is possible in areas where FLAER may not be widely available. © 2016 International Clinical Cytometry Society.
Dahl Chase Diagnostic Services Bangor Maine
Department of Laboratory Medicine Toronto General Hospital Toronto Ontario Canada
Exbio Praha Prague Czech Republic
Institute of Hematology and Blood Transfusion Prague Czech Republic
References provided by Crossref.org
