HLA-B-ASTERISK-27 TYPING BY GROUP-SPECIFIC HYBRIDIZATION IN MICROTITER PLATES

We have developed and evaluated a test for HLA-B27 based on PCR and D NA hybridization in microtiter plates. A region within exon 2 of the H LA-B gene is amplified and labeled by PCR and the amplification produc t is hybridized to a group-specific HLA-B27 and a generic control oli gonucleotide probe in two separate cavities of the plate. Bound sequen ces are detected using an ELISA-like protocol. The assay has been eval uated on 254 DNA samples routinely received for B27 testing in paralle l with serological and SSP-PCR typing Results were concordant in typin g 102 HLA-B27-positive and 152 HLA-B27-negative individuals except for two samples containing HLA-B73, which stained B27 positive in the mi crowell test. The new procedure is rapid and simple to perform, and th e microwell format is particularly suitable for automation.