Flow Cytometric Human Leukocyte Antigen-B27 Typing with Stored Samples for Batch Testing
Annals of Laboratory Medicine
; : 174-183, 2013.
Article
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| ID: wpr-144110
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WPRO
ABSTRACT
BACKGROUND: Flow cytometry (FC) HLA-B27 typing is still used extensively for the diagnosis of spondyloarthropathies. If patient blood samples are stored for a prolonged duration, this testing can be performed in a batch manner, and in-house cellular controls could easily be procured. In this study, we investigated various methods of storing patient blood samples. METHODS: We compared four storage methods: three methods of analyzing lymphocytes (whole blood stored at room temperature, frozen mononuclear cells, and frozen white blood cells [WBCs] after lysing red blood cells [RBCs]), and one method using frozen platelets (FPLT). We used three ratios associated with mean fluorescence intensities (MFI) for HLAB27 assignment: the B27 MFI ratio (sample/control) for HLA-B27 fluorescein-5-isothiocyanate (FITC); the B7 MFI ratio for HLA-B7 phycoerythrin (PE); and the ratio of these two ratios, B7/B27 ratio. RESULTS: Comparing the B27 MFI ratios of each storage method for the HLA-B27+ samples and the B7/B27 ratios for the HLA-B7+ samples revealed that FPLT was the best of the four methods. FPLT had a sensitivity of 100% and a specificity of 99.3% for HLA-B27 assignment in DNA-typed samples (N=164) when the two criteria, namely, B27 MFI ratio >4.0 and B7/B27 ratio <1.5, were used. CONCLUSIONS: The FPLT method was found to offer a simple, economical, and accurate method of FC HLA-B27 typing by using stored patient samples. If stored samples are used, this method has the potential to replace the standard FC typing method when used in combination with a complementary DNA-based method.
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Índice:
WPRIM
Asunto principal:
Temperatura
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Plaquetas
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Prueba de Histocompatibilidad
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Leucocitos Mononucleares
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Antígeno HLA-B7
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Antígeno HLA-B27
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Espondiloartropatías
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Eritrocitos
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Reacción en Cadena en Tiempo Real de la Polimerasa
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Citometría de Flujo
Límite:
Humans
Idioma:
En
Revista:
Annals of Laboratory Medicine
Año:
2013
Tipo del documento:
Article