Rapid typing of DNA sequence polymorphism at the HLA-DRB1 locus using the polymerase chain reaction and nonradioactive oligonucleotide probes.
An HLA-DR typing system that uses sequence-specific oligonucleotide (SSO) probes conjugated to horseradish peroxidase (HRP) for analyzing DRB alleles amplified by the polymerase chain reaction has been developed. Using 25 HRP-SSO probes and two primer pairs for generic and for DRB1 locus-specific amplification, we can distinguish 31 of 34 HLA-DRB1 alleles. This procedure is suitable for typing heterozygous samples from a variety of sources, including cDNA templates, and can detect in a simple and rapid dot-blot format allelic variants not distinguishable by serological methods. It should prove valuable for tissue typing, determining individual identity, and studies of disease susceptibility.
Scharf
S J
SJ
Department of Human Genetics, Cetus Corporation, Emeryville, California 94608.
Griffith
R L
RL
Erlich
H A
HA
D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA / Q000235:genetics*; D015342:DNA Probes; D005091:Exons; D006684:HLA-DR Antigens / Q000145:classification / Q000235:genetics*; D059811:HLA-DRB1 Chains; D000949:Histocompatibility Antigens Class II / Q000145:classification / Q000235:genetics*; D006650:Histocompatibility Testing; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic
D015342:DNA Probes; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains; D000949:Histocompatibility Antigens Class II; D004247:DNA