title: HLA-DPB1 typing by PCR-SSO reverse dot blot hybridization after group-specific amplification.
abstract: 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.
authors.affiliation: Blutbank, Abteilung für Innere Medizin mit Schwerpunkt Hämatologie und Onkologie, Universitätsklinikum Rudolf Virchow, Freie Universität, Berlin.
authors.forename: R
authors.initials: R
authors.lastname: Blasczyk
authors.forename: M
authors.initials: M
authors.lastname: Mohr
authors.forename: R
authors.initials: R
authors.lastname: Zimmermann
authors.forename: N
authors.initials: N
authors.lastname: Schwella
authors.forename: D
authors.initials: D
authors.lastname: Huhn
mesh_terms: 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
chemical_list: D006682:HLA-DP Antigens; D059868:HLA-DP beta-Chains; C071170:HLA-DPB1 antigen; D015345:Oligonucleotide Probes