Clinical relevance of CD157 for rapid and cost-effective simultaneous evaluation of PNH granulocytes and monocytes by flow cytometry

• Published in: International journal of laboratory hematology 2015-04, Vol.37 (2), p.231-237
• Main Authors: Marinov, I., Kohoutová, M., Tkáčová, V., Pešek, A., Čermák, J., Cetkovský, P.
• Format: Article
• Language: English
• Subjects: ADP-ribosyl Cyclase - metabolism; Antigens, CD - metabolism; Biomarkers; CD157; clinical relevance; flow cytometry; Flow Cytometry - methods; Flow Cytometry - standards; GPI-Linked Proteins - metabolism; Granulocytes - metabolism; Hemoglobinuria, Paroxysmal - diagnosis; Hemoglobinuria, Paroxysmal - metabolism; Humans; Immunophenotyping - methods; Immunophenotyping - standards; Leukocytes - metabolism; Monocytes - metabolism; Paroxysmal nocturnal hemoglobinuria; Reproducibility of Results
• ISSN: 1751-5521; 1751-553X
• DOI: 10.1111/ijlh.12271

Summary Introduction CD157 has been reported as a potentially useful marker for paroxysmal nocturnal hemoglobinuria (PNH) testing by flow cytometry (FCM). Methods We determined the performance characteristics of a CD157‐based five‐color assay and compared results from patient analysis with results obtained with a previously validated CD14/CD24‐based six‐color protocol. Results Coefficient of variation (CV) for intra‐/interassay precision analysis of granulocytes ranged from 0.88/0.09% to 3.02/3.71% and from 0.20/0.08% to 8.83/4.04% for the five‐ and six‐color protocol, respectively. For monocyte, CV ranged from 0.42/0.49% to 8.13/4.80% for the five‐color protocol and from 0.28/0.70% to 5.41/3.19% for the six‐color protocol within various PNH clones. Coefficient of determination (r2) for linear regression analysis of PNH clones ranging from 0.3 to 99.8% was >0.99 in all cases, Wilcoxon ranks test showed no statistically significant differences (P > 0.05), and Bland–Altman analysis demonstrated agreement with mean bias ranging from −0.02 to 0.38. Conclusion Our results confirm very good performance characteristics for both intra‐ and interassay precision analyses, excellent correlation, and agreement between approaches. In agreement with recently published data, our experience supports the clinical relevance of CD157 for a rapid and cost‐effective simultaneous evaluation of PNH leukocytes by flow cytometry.