Long-range PCR amplification as an alternative strategy for characterizing novel HLA-B alleles.
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.
Curran
M D
MD
Northern Ireland Tissue Typing Laboratory, City Hospital, Belfast, Northern Ireland, UK.
Williams
F
F
Earle
J A
JA
Rima
B K
BK
van Dam
M G
MG
Bunce
M
M
Middleton
D
D
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
D015235:HLA-B Antigens; D015797:HLA-B35 Antigen; C056322:HLA-B41 antigen; D004247:DNA