Louis Waeckel, Anne Kennel, Anne-Emmanuelle Berger, Claude Lambert
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引用次数: 1
Abstract
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-B27TM 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.
期刊介绍：
Cytometry Part B: Clinical Cytometry features original research reports, in-depth reviews and special issues that directly relate to and palpably impact clinical flow, mass and image-based cytometry. These may include clinical and translational investigations important in the diagnostic, prognostic and therapeutic management of patients. Thus, we welcome research papers from various disciplines related [but not limited to] hematopathologists, hematologists, immunologists and cell biologists with clinically relevant and innovative studies investigating individual-cell analytics and/or separations. In addition to the types of papers indicated above, we also welcome Letters to the Editor, describing case reports or important medical or technical topics relevant to our readership without the length and depth of a full original report.
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