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HLA-B27 detection - comparison of genetic sequence-based method and flow cytometry assay
Urszula Skalska1, Anna Kozakiewicz1, Włodzimierz Maśliński1
1Department of Pathophysiology and Immunology, Institute of Rheumatology, Warsaw, Poland.
Reumatologia
|July 14, 2016
Summary
Human Leukocyte Antigen B27 (HLA-B27) testing is crucial for diagnosing ankylosing spondylitis. A genetic sequence-based method and flow cytometry assay show 99% agreement, with HLA-B27 subtype 27:05 being most common.
Area of Science:
- Immunogenetics
- Molecular Diagnostics
Background:
- Human Leukocyte Antigen B27 (HLA-B27) is a key genetic marker strongly associated with ankylosing spondylitis.
- Routine HLA-B27 testing aids in the diagnosis of spondyloarthropathies.
Purpose of the Study:
- To compare the diagnostic accuracy of a genetic sequence-based method versus a flow cytometry assay for HLA-B27 detection.
- To identify prevalent HLA-B27 allelic variations in individuals with suspected spondyloarthropathy.
Main Methods:
- Peripheral blood samples from 300 individuals were analyzed.
- Flow cytometry assay using GS145.2 monoclonal antibody for HLA-B27 expression.
- DNA analysis via polymerase chain reaction (PCR) and gene sequencing to detect HLA-B27 alleles.
Main Results:
- Flow cytometry identified 76 HLA-B27 positive samples; genetic sequencing confirmed 73.
- A high concordance rate of 99% was observed between the two methods.
- The most frequent HLA-B27 allelic variation identified was 27:05, followed by 27:02 and 27:07.
Conclusions:
- Genetic sequencing and flow cytometry are highly consistent methods for HLA-B27 detection.
- The prevalence of HLA-B27 subtype 27:05 highlights its significant association with ankylosing spondylitis risk.
