Oligonucleotide probing. Applications to HLA typing.
Human leukocyte antigen typing is currently being done by both serologic and cellular techniques. It is this methodology that forms the basis for organ transplantation, since HLA antigens are the major proteins involved in graft rejection. Matching organ graft donors with recipients at these polymorphic HLA loci clearly increases graft survival in most transplant systems. However, the serologic and cellular techniques have shortcomings, including the problem of standardizing antisera, the need for technologists with expertise in T-cell cloning, and the failure to work successfully with some cells (eg, leukemic blasts). We have begun to develop a more rapid, sensitive typing method using molecular biological techniques to tissue-type cells. Using oligonucleotide probes, we can now identify HLA-D region alleles (which may differ by only a single nucleotide) that can only be differentiated by a highly technical, lengthy cellular technique. Oligonucleotide probing not only provides the basis for a rapid method of HLA typing, which in all likelihood will become generally applicable as additional sequence information becomes available, but also allows definition of a gene transcript from an individual locus rather than the phenotypic results provided by current HLA typing techniques.
Freeman
S M
SM
Department of Laboratory Medicine/Pathology, University of Minnesota, Minneapolis.
Noreen
H J
HJ
Bach
F H
FH
D000483:Alleles; D006680:HLA Antigens / Q000145:classification* / Q000235:genetics; D006684:HLA-DR Antigens / Q000145:classification / Q000235:genetics; D015804:HLA-DR4 Antigen; D006239:Haplotypes; D006801:Humans; D008967:Molecular Biology / Q000379:methods; D009841:Oligonucleotides; D014158:Transcription, Genetic
D006680:HLA Antigens; D006684:HLA-DR Antigens; D015804:HLA-DR4 Antigen; D009841:Oligonucleotides