Identification of Human Leukocyte Antigens, HLA, for matching of kidney and bone marrow donors and recipients is mandatory for transplantation. Many allele combinations fail to be resolved with current typing methods. Haplotype-Specific Extraction, HSE, establishes haplotypes from an individual patient's sample, and permits the unambiguous typing without knowledge of familial information. It is the goal of the proposed work to develop HSE into: 1) a reliable method to improve HLA typing. 2) a powerful tool for researchers studying the implications of HLA and disease. During Phase I, HSE was developed for HLA-B. There are currently 498 HLA-B alleles identified with 1068 ambiguous allele combinations. The use of HSE for separation of HLA-B alleles into haploid components was successfully demonstrated in nine HLA laboratories. A preliminary product for separation of common ambiguous combinations of HLA-B alleles was produced, and HSE was automated on a GenoM-6 robot, for consistent, walk-away processing for one to six haploid extractions. Under the proposed Phase II funding HSE will be expanded for use at other HLA loci, other HLA-linked genes, and polymorphic micro-satellite markers in the Major Histocompatibility Complex, MHC. It is a further goal of Phase II to demonstrate the direct compatibility of HSE with existing SNP-mapping information and to expand the automation of HSE to a 96-well platform.
