HLA‐DPB typing using co‐digestion of amplified fragments allows efficient identification of heterozygous genotypes

Abstract

Twenty-four alleles have been defined for HLA-DPB based on their second exon sequences. This paper describes a novel method, co-digested amplified fragment length polymorphisms (CAFLP), for assigning these alleles to heterozygous patients, as well as to homozygous cell lines. The method depends on co-digestion of amplified DNA by restriction endonucleases and separation of the resultant fragments with polyacrylamide gel electrophoresis. Co-digestion by selected restriction enzymes produces a set of readily discernible fragments that are unique for a given haplotype because the selected restriction sites occur in cis. Consequently, this method provides haplotype information not available from independent digests and allows all known heterozygous genotypes to be identified. Analysis of 103 trios of mother, father, and child, plus 120 normal caucasians, demonstrates the reliability and simplicity of this procedure. This simple typing method results in unambiguous assignment of all current HLA-DPB genotypes in random samples with a high proportion of heterozygous individuals.