HLA-C HIGH-RESOLUTION TYPING - ANALYSIS OF EXON-2 AND EXON-3 BY SEQUENCE BASED TYPING AND DETECTION OF POLYMORPHISMS IN EXONS 1-5 BY SEQUENCE-SPECIFIC PRIMERS

HLA-C high resolution sequence based typing developed in this study in volves a unique DNA amplification encompassing exon 1 to intron 3 and four fluorescent sequencing reactions covering exon 2 and 3. Both dye primer and dye terminator sequencing techniques were performed and res ults compared. This approach allowed the identification of all of the 50 HLA-C allelic variants so far described, except for two allele pair s that are distinguished by non-coding nucleotide changes (Cw12021=12 022, Cw15051 = 15052) and three allele pairs (Cw*0701 = 0706, Cw*1701 = 1702 and Cw1801 = 1802) that share the same nucleotide sequence in exon 2 and 3. For complete subtyping of these allelic variants, an am plification based on sequence specific primers (PCR-SSP) was used. No ambiguous heterozygous combinations of alleles were detected in our pa nel so far. HLA-C typing data obtained by this method were compared wi th data from serological and low resolution PCR-SSP typing, which had been performed previously on the samples sequenced.