HLA-DPB1 typing by PCR-SSO reverse dot blot hybridization after group-specific amplification.
BACKGROUND
The allelic diversity of HLA-DPB1 antigens can be determined at the DNA level after PCR amplification. The pattern of polymorphism at the DPB1 locus makes it difficult to unambiguously assign all genotypes in a typing system using one single pair of generic primers.


METHODS
We apply here a simple technique based on the reverse dot blot analysis to the typing of HLA-DPB1 alleles. In order to increase its resolution, a group-specific amplification based on sequence variations of the polymorphic region F was used subdividing the HLA-DPB1 alleles in 2 nonoverlapping families. A separate analysis was then performed within each group of alleles.


RESULTS
Using these 2 primer pairs, 21 group 1 and 30 group 2 alleles were separately amplified. From 1,378 possible allele combinations for DPB1*0101-5301 only 33 gave ambiguous typing results compared to 61 using a single pair of generic primers.


CONCLUSIONS
This procedure provides a rapid and simple HLA-DPB1 genotyping. Especially in heterozygotes the hybridization patterns were easier to interpret. The utilization of group-specific amplification substantially reduced ambiguous typing results.
Blasczyk
R
R
Blutbank, Abteilung für Innere Medizin mit Schwerpunkt Hämatologie und Onkologie, Universitätsklinikum Rudolf Virchow, Freie Universität, Berlin.
Mohr
M
M
Zimmermann
R
R
Schwella
N
N
Huhn
D
D
D000483:Alleles; D016026:Bone Marrow Transplantation; D005784:Gene Amplification / Q000235:genetics*; D005838:Genotype; D006682:HLA-DP Antigens / Q000235:genetics*; D059868:HLA-DP beta-Chains; D006579:Heterozygote; D006650:Histocompatibility Testing; D006801:Humans; D015151:Immunoblotting; D007938:Leukemia / Q000235:genetics; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic
D006682:HLA-DP Antigens; D059868:HLA-DP beta-Chains; C071170:HLA-DPB1 antigen; D015345:Oligonucleotide Probes