title: HLA-DQA1 allele and suballele typing using noncoding sequence polymorphisms. Application to 4AOHW cell panel typing.
abstract: HLA-DQA1 typing of the 4AOHW cell panel is presented using a novel strategy that exploits both intron and exon polymorphisms. Intron sequences adjacent to the variable HLA-DQA1 second exon exhibit stable polymorphisms that are specific for locus alleles and certain suballelic DR/DQ haplotypes. A PCR-RFLP method has been developed that is based on amplification of a 780-bp segment extending from intron 1 through exon 2 to intron 2. Stable sequence polymorphisms provide restriction enzyme sites and confer mobility variations detected on polyacrylamide minigel electrophoresis. Direct band comparison of amplified products and restriction fragments with known standards facilitates pattern comparison, obviating the requirement for accurate molecular weight determination. This method, using only two enzymes, identifies a total of 11 allelic and suballelic groups, including all eight DQA1 alleles encoded at the second exon.
authors.affiliation: GeneType Pty Ltd., Fitzroy, Australia.
authors.forename: T M
authors.initials: TM
authors.lastname: Limm
authors.forename: M L
authors.initials: ML
authors.lastname: Ashdown
authors.forename: M J
authors.initials: MJ
authors.lastname: Naughton
authors.forename: M D
authors.initials: MD
authors.lastname: McGinnis
authors.forename: M J
authors.initials: MJ
authors.lastname: Simons
mesh_terms: D000483:Alleles; D001483:Base Sequence; D002461:Cell Line, Transformed / Q000276:immunology; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005091:Exons / Q000235:genetics*; D006683:HLA-DQ Antigens / Q000145:classification / Q000235:genetics*; D059848:HLA-DQ alpha-Chains; D006684:HLA-DR Antigens; D006239:Haplotypes; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007438:Introns / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
chemical_list: D006683:HLA-DQ Antigens; D059848:HLA-DQ alpha-Chains; C069050:HLA-DQA1 antigen; D006684:HLA-DR Antigens; C040837:endodeoxyribonuclease MaeII; D015252:Deoxyribonucleases, Type II Site-Specific; C040158:GTAC-specific type II deoxyribonucleases