Flow Cytometric Human Leukocyte Antigen-B27 Typing with Stored Samples for Batch Testing
Annals of Laboratory Medicine
;
: 174-183, 2013.
Article
in English
| WPRIM
| ID: wpr-144110
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 storagemethods:
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Temperature
/
Blood Platelets
/
Histocompatibility Testing
/
Leukocytes, Mononuclear
/
HLA-B7 Antigen
/
HLA-B27 Antigen
/
Spondylarthropathies
/
Erythrocytes
/
Real-Time Polymerase Chain Reaction
/
Flow Cytometry
Limits:
Humans
Language:
English
Journal:
Annals of Laboratory Medicine
Year:
2013
Type:
Article
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