NONRADIOISOTOPIC TYPING OF HUMAN-LEUKOCYTE ANTIGEN CLASS-II GENES ON MICROPLATES

We describe a new nonradioisotopic method for HLA class II molecular t yping performed in 96-well plates of the same size as those used in en zyme-linked immunosorbent assays (ELISA) and radioimmunoassays (RIA). Biotinylated sequence-specific oligonucleotide (SSO) probes are bound to avidin-coated plates. Digoxigenin-labeled PCR-amplified DNA samples are hybridized, washed and detected with a peroxidase conjugated anti body assay. The method was tested by performing a partial HLA DQA1 and DQB1 typing on 69 randomly selected blood samples The results are com pletely concordant with a traditional SSO-PCR typing performed on the same samples. This procedure is simple, fast and could be adapted for performance in semi-automated or automated fashion using equipment alr eady available for ELISA and RIA assays.