HLA-B*27 typing by group-specific hybridization in microtiter plates.
We have developed and evaluated a test for HLA-B*27 based on PCR and DNA hybridization in microtiter plates. A region within exon 2 of the HLA-B gene is amplified and labeled by PCR and the amplification product is hybridized to a group-specific HLA-B*27 and a generic control oligonucleotide probe in two separate cavities of the plate. Bound sequences are detected using an ELISA-like protocol. The assay has been evaluated on 254 DNA samples routinely received for B27 testing in parallel with serological and SSP-PCR typing. Results were concordant in typing 102 HLA-B27-positive and 152 HLA-B27-negative individuals except for two samples containing HLA-B*73, which stained B27 positive in the microwell test. The new procedure is rapid and simple to perform, and the microwell format is particularly suitable for automation.
Department of Transfusion Medicine, University of Graz, Austria. thomas.wagner@kfunigraz.ac.at
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Wagner
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Oberkanins
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Weinmayr
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Helmberg
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Kury
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Lanzer
D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation / Q000379:methods*; D015796:HLA-B27 Antigen / Q000032:analysis* / Q000235:genetics*; D015395:Histocompatibility Antigens Class I / Q000235:genetics; D006801:Humans; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D012371:Robotics
D015796:HLA-B27 Antigen; D015395:Histocompatibility Antigens Class I; D015345:Oligonucleotide Probes