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HLA-B*27 typing by group-specific hybridization in microtiter plates
T Wagner1, C Oberkanins, B Weinmayr
1Department of Transfusion Medicine, University of Graz, Austria. thomas.wagner@kfunigraz.ac.at
Tissue Antigens
|October 2, 1998
Summary
A new HLA-B*27 DNA test using PCR and microtiter plate hybridization offers a rapid and automatable method for identifying HLA-B27 status. This test shows high concordance with existing methods, improving diagnostic efficiency.
Area of Science:
- Molecular Biology
- Immunogenetics
- Clinical Diagnostics
Background:
- The human leukocyte antigen B*27 (HLA-B*27) is strongly associated with seronegative spondyloarthropathies.
- Accurate and efficient HLA-B*27 typing is crucial for diagnosis and patient management.
Purpose of the Study:
- To develop and validate a novel, rapid, and automatable DNA-based test for HLA-B*27 detection.
- To evaluate the performance of the new test against established serological and SSP-PCR methods.
Main Methods:
- Polymerase chain reaction (PCR) amplification and labeling of a specific region within exon 2 of the HLA-B gene.
- Hybridization of the amplified product to group-specific HLA-B*27 and generic control oligonucleotide probes in a microtiter plate format.
- Detection of bound sequences using an enzyme-linked immunosorbent assay (ELISA)-like protocol.
Main Results:
- The assay was evaluated on 254 DNA samples, correctly identifying 102 HLA-B*27-positive and 152 HLA-B*27-negative individuals.
- High concordance was observed when compared to serological and SSP-PCR typing methods.
- Two samples with HLA-B*73 showed a false-positive result for HLA-B*27 in the microwell test.
Conclusions:
- The developed PCR and DNA hybridization assay provides a fast, simple, and automatable method for HLA-B*27 testing.
- The microtiter plate format is well-suited for high-throughput clinical laboratories.
- While generally accurate, potential cross-reactivity with HLA-B*73 requires consideration.