title: Short-form HLA-DP typing with 48 primer mixes using the polymerase chain reaction and sequence-specific primers.
abstract: We have developed a short-form SSP-based HLA-DP typing system for routine use adapted from a comprehensive HLA-DP typing method described by Gilchrist et at. (1998). Our short-form system detects 93 alleles, including the 18 most common HLA-DPB1 alleles and eight HLA-DPA1 alleles. The primer mixes described were tested using the PCR-SSP Manager (Bunce et al., 1998) database to confirm the specificity of selected primers, and to detect potentially ambiguous amplifications. This short-form HLA-DP typing system was validated using 50 fully typed samples obtained through the UCLA International DNA Exchange. All samples gave 100% concordance with the consensus type. Our laboratory routinely uses a PCR-SSP based system of 48 primer mixes for HLA-DRB and HLA-DQB typing. The advantage of the short-form HLA-DP typing system described here is that the additional 48 HLA-DP primer mixes required can be included on the second half of a 96-well format tray. This method now enables a full HLA class II typing at the level of allele group resolution in 2 1/2 h.
authors.affiliation: Transplantation Laboratory, Manchester Institute of Nephrology and Transplantation, Manchester Royal Infirmary, UK.
authors.forename: J A
authors.initials: JA
authors.lastname: Davidson
authors.forename: K V
authors.initials: KV
authors.lastname: Poulton
mesh_terms: D000483:Alleles; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D006682:HLA-DP Antigens / Q000235:genetics*; D059867:HLA-DP alpha-Chains; D059868:HLA-DP beta-Chains; D006239:Haplotypes; D006650:Histocompatibility Testing / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D006682:HLA-DP Antigens; D059867:HLA-DP alpha-Chains; D059868:HLA-DP beta-Chains; C071172:HLA-DPA1 antigen; C071170:HLA-DPB1 antigen