Defining the allelic variants of HLA-A30 in the Sardinian population using amplification refractory mutation system--polymerase chain reaction.
HLA-A30 is present in the Sardinian population at a frequency of 23%. We have designed a system using nested ARMS-PCR to determine the relative frequencies of the HLA-A*30 allelic variants (A*3001, A*3002, and A*3003) within this population. The use of a nested PCR approach, in which the first-round reaction provides HLA-A*30 specificity and template DNA for the subsequent nested reactions, is a powerful means of discriminating between alleles of very similar sequence. Using this method, we performed subtyping of 35 serologically defined HLA-A30 Sardinian individuals, and taking into account homozygotes, identified 38 A*30 alleles. Of these, 33 typed as A*3002, four typed as A*3001, and one sample did not conform to the patterns of reactivity of any of the published A*30 alleles. Haplotype information showed strong linkage disequilibrium between A*3002 and B18. This study underlines the potential of DNA-based methods for typing HLA class I in terms of adding further levels of definition to studies of population structure and also as a means of identifying new alleles.
Krausa
P
P
ICRF, Institute of Molecular Medicine, Oxford, UK.
Carcassi
C
C
Orrù
S
S
Bodmer
J G
JG
Browning
M J
MJ
Contu
L
L
D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D016384:Consensus Sequence; D004252:DNA Mutational Analysis; D005787:Gene Frequency; D014644:Genetic Variation; D005838:Genotype; D015234:HLA-A Antigens / Q000235:genetics*; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing; D007558:Italy / Q000453:epidemiology; D015810:Linkage Disequilibrium; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D015234:HLA-A Antigens; C069526:HLA-A*30 antigen