DIRECT SEQUENCING OF SSP-PCR-AMPLIFIED CDNA TO IDENTIFY NEW ALLELES IN THE DR52-ASSOCIATED DRB1 GROUP - IDENTIFICATION OF DRB1-ASTERISK-1115, DRB1-ASTERISK-1117 AND DRB1-ASTERISK-1319

Low and high resolution sequence specific oligonucleotide probe hybrid ization patterns were used to design an approach to direct sequencing of allele specific amplified cDNA. Several PCR amplifications were use d to derive overlapping sequence fragments to define complete first do main sequences for a single allele. This method has been used to chara cterize three new DRB1 alleles in the DR52 family, DRB11115, DRB1*111 7, and DRB11319. AU three alleles carry polymorphisms previously obse rved in other DRB alleles and underscore the importance of utilizing a directed sequencing approach for obtaining unambiguous typing results in matching for bone marrow transplantation between unrelated donor a nd recipient.