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.
Wagner
T
T
Department of Transfusion Medicine, University of Graz, Austria. thomas.wagner@kfunigraz.ac.at
Oberkanins
C
C
Weinmayr
B
B
Helmberg
W
W
Kury
F
F
Lanzer
G
G
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