title: Long-range PCR amplification as an alternative strategy for characterizing novel HLA-B alleles.
abstract: We have developed a simple, rapid and reliable method for specifically amplifying and cloning full-length HLA-B genes from genomic DNA. Using this methodology we characterized three alleles of interest at the molecular level. Two of the alleles appeared in our routine class I PCR-SSOP typing system, a variant of B*5801 found in the Daudi cell line and RCE 56 and a variant of B*4101 found in a number of volunteer donors on our Bone Marrow Donor Registry. The third, a variant B35 allele found in RCE 80, was first identified as unusual by serology. Our sequencing analysis of exon 2 and exon 3 identified two of these alleles as the recently reported novel HLA-B*5802 and HLA-B*4102 alleles, while the third represents a new B35 allele officially designated B*3513.
authors.affiliation: Northern Ireland Tissue Typing Laboratory, City Hospital, Belfast, Northern Ireland, UK.
authors.forename: M D
authors.initials: MD
authors.lastname: Curran
authors.forename: F
authors.initials: F
authors.lastname: Williams
authors.forename: J A
authors.initials: JA
authors.lastname: Earle
authors.forename: B K
authors.initials: BK
authors.lastname: Rima
authors.forename: M G
authors.initials: MG
authors.lastname: van Dam
authors.forename: M
authors.initials: M
authors.lastname: Bunce
authors.forename: D
authors.initials: D
authors.lastname: Middleton
mesh_terms: D000483:Alleles; D001483:Base Sequence; D004247:DNA; D014644:Genetic Variation; D015235:HLA-B Antigens / Q000235:genetics*; D015797:HLA-B35 Antigen / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured
chemical_list: D015235:HLA-B Antigens; D015797:HLA-B35 Antigen; C056322:HLA-B41 antigen; D004247:DNA