1. Evaluation of paroxysmal nocturnal hemoglobinuria screening by flow cytometry through multicentric interlaboratory comparison in four countries.
- Author
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Debliquis, Agathe, Wagner-Ballon, Orianne, Le Garff-Tavernier, Magali, Fossat, Chantal, Chatelain, Bernard, Letestu, Rémi, Drénou, Bernard, and HPN-AFC Group
- Subjects
PAROXYSMAL hemoglobinuria ,FLOW cytometry ,AEROLYSIN ,ROBUST statistics ,BLOOD sampling ,HEMOLYTIC anemia diagnosis ,COMPARATIVE studies ,DIAGNOSTIC errors ,RESEARCH methodology ,MEDICAL cooperation ,RESEARCH ,EVALUATION research ,STANDARDS - Abstract
Objectives: Paroxysmal nocturnal hemoglobinuria (PNH) is currently diagnosed by flow cytometry; although highly sensitive, its interpretation and reporting appear as critical as its technique. Thus, we developed a quality control scheme for the French-speaking region based on the international recommendations for PNH screening.Methods: After a topical workshop, we proposed a 1-year, two-step survey program to any volunteering French-speaking clinical laboratory. The first survey consisted of sending raw data files to evaluate gating and the interpretation strategy of each center. The second stipulated sending fresh whole-blood samples to evaluate the whole process and its practice.Results: Forty-nine participants from voluntary centers returned results for each of the two successive surveys. On virtual survey, 27% reported false-positive PNH created by immature granulocytes, whereas the minor PNH clone was not detected by 9%. On fresh survey, 63% of centers used at least the same six-color combination (CD24, CD14, CD33, CD15, CD45, and fluorescent aerolysin), and nearly 70% of participants were able to perform a sensitivity test less than 0.1% on neutrophils. All participants detected the major PNH clone, yet 16% returned false-positive results for the non-PNH clone case.Conclusions: We succeeded in rallying numerous French-speaking clinical laboratories for both surveys and in harmonizing the technical practice by highlighting common pitfalls. [ABSTRACT FROM AUTHOR]- Published
- 2015
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