High resolution definition of HLA-DRB haplotypes by a simplified microsatellite typing technique.
In humans, the region configurations DR1, DR8, DR51, DR52 and DR53 are known to display copy number as well as allelic variation, rendering high resolution typing of HLA-DRB haplotypes cumbersome. Advantage was taken of microsatellite D6S2878, present in all DRB genes/pseudogenes with an intact exon 2-intron 2 segment. This DRB-STR is highly polymorphic in composition and length. Recently, it was proven that all exon 2 sequences could be linked to a certain DRB-STR that segregates with the respective DRB allele. Because haplotypes show differential copy numbers and compositions of exon 2-positive DRB genes/pseudogenes, unique DRB-STR patterns could be described that appear to be specific for a particular DRB haplotype. The aim of this workshop project was to approve and to qualify this simple typing protocol in a larger panel covering different European populations. All participants succeeded in correctly defining the DRB-STR amplicons varying from 135 to 222 base pair (bp) lengths. The panel of 101 samples covered 50 DRB alleles distributed over 37 different haplotypes as defined by exon 2 sequence-based typing. These haplotypes could be refined into 105 haplotypes by DRB-STR typing. Thus, discrimination of exon 2-identical DRB alleles was feasible, as well as the exact description of three different crossing-over events that resulted in the generation of hybrid DR region configurations. This typing procedure appears to be a quick and highly robust technique that can easily be performed by different laboratories, even without experience in microsatellite typing; thus, it is suitable for a variety of researchers in diverse research areas.
Department of Comparative Genetics & Refinement, Biomedical Primate Research Centre, Rijswijk, The Netherlands. doxiadis@bprc.nl
G G M
GG
Doxiadis
N
N
de Groot
E-M
EM
Dauber
P H
PH
van Eede
I
I
Fae
R
R
Faner
G
G
Fischer
Z
Z
Grubic
N M
NM
Lardy
W
W
Mayr
E
E
Palou
W
W
Swelsen
K
K
Stingl
I I N
II
Doxiadis
R E
RE
Bontrop
D000818:Animals; D019143:Evolution, Molecular; D006684:HLA-DR Antigens / Q000235:genetics*; D006239:Haplotypes; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics*
D006684:HLA-DR Antigens