Human leukocyte antigen-B27 typing by flow cytometry: Comparison of three CE-IVD methods.

Autor: Waeckel L; Laboratoire d'immunologie clinique, Hôpital Nord, Saint-Étienne, France., Kennel A; Établissement Français du Sang Auvergne-Rhône-Alpes, Saint-Étienne, France., Berger AE; Laboratoire d'immunologie clinique, Hôpital Nord, Saint-Étienne, France., Lambert C; Laboratoire d'immunologie clinique, Hôpital Nord, Saint-Étienne, France.
Jazyk: angličtina
Zdroj: Cytometry. Part B, Clinical cytometry [Cytometry B Clin Cytom] 2023 Nov; Vol. 104 (6), pp. 460-467. Date of Electronic Publication: 2022 Nov 30.
DOI: 10.1002/cyto.b.22102
Abstrakt: Background: The human leukocyte antigen B27 (HLA-B27), associated with chronic inflammatory diseases is thus widely performed for diagnostic purposes. Genotyping by molecular biology is the current gold standard for HLA-B27 typing, but cheaper and faster flow cytometry methods have been developed.
Methods: In this report, we compare analytical performances of three CE-IVD flow cytometry kits: IOTest™ and DuraClone B27™ from Beckman Coulter and BD HLA-B27™ from BD Biosciences on a Navios™ cytometer as compared to molecular biology.
Results: Routine analyses could conclude for HLA-B27 in197 patients (23.2%) and was not conclusive for 66 patients (7%, 8%). The experience revealed the needs to complete IOTest™ with a lineage marker (like CD3-APC) and a standardization procedure in detection of fluorescence. Comparative analysis considering 23/44 non-conclusive samples as negative, pointed out a 100% sensitivity and specificity of IOTest™ in determining genetically proved HLA-B27. Specificity of the BD HLA-B27 TM kit was 94% (two false positives) sticking to the manufacturer instructions but raised to 100% and 94% sensitivity with a cutoff at 8.5 (225 on FACSdiva's scale).
Discussion: The DuraClone B27™ specificity was poor using the manufacturer cutoff but raised to 100% with a 8.0 cutoff instead of 15.
Conclusion: The three flow cytometry kits displayed satisfying performances but need adjustments. The DuraClone B27™ kit seems to be the best while the IOTest™ kit is not conclusive in 8% of cases. In most cases the use of molecular biology can be spared, but genotyping remains essential for patients whose HLA-B27 status cannot be determined with confidence by flow cytometry.
(© 2022 The Authors. Cytometry Part B: Clinical Cytometry published by Wiley Periodicals LLC on behalf of International Clinical Cytometry Society.)
Databáze: MEDLINE
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