title: HLA-DQB1 and DQA1 matching by ambient temperature PCR-SSCP.
abstract: We have developed a polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) protocol for rapid matching of DQA1 and DQB1 alleles. Electrophoresis can be performed at ambient temperature within the range 18-28 degrees C without continuous gel cooling. The method has been tested on 27 patient-potential bone marrow donor pairs for DQB1 and 31 pairs for DQA1. Bone marrow pairs were chosen to represent a broad range of common alleles based upon previous restriction fragment length polymorphism (RFLP) analysis type assignments. Samples were re-typed by PCR with sequence-specific primers (PCR-SSP) and the results compared to matching by PCR-SSCP analysis. There was a 100% correlation between PCR-SSP and PCR-SSCP analysis for DQB1, and a 97% correlation for DQA1 matching.
authors.affiliation: Department of Transplantation Sciences, University of Bristol, UK.
authors.forename: T M
authors.initials: TM
authors.lastname: Clay
authors.forename: D
authors.initials: D
authors.lastname: Culpan
authors.forename: M C
authors.initials: MC
authors.lastname: Pursall
authors.forename: B A
authors.initials: BA
authors.lastname: Bradley
authors.forename: J L
authors.initials: JL
authors.lastname: Bidwell
mesh_terms: D001483:Base Sequence; D017931:DNA Primers; D005069:Evaluation Studies as Topic; D006683:HLA-DQ Antigens / Q000235:genetics*; D059848:HLA-DQ alpha-Chains; D059866:HLA-DQ beta-Chains; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D018807:Polymorphism, Single-Stranded Conformational; D015203:Reproducibility of Results; D013696:Temperature
chemical_list: D017931:DNA Primers; D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; D059866:HLA-DQ beta-Chains; C069050:HLA-DQA1 antigen; C069051:HLA-DQB1 antigen