Direct sequencing of SSP-PCR-amplified cDNA to identify new alleles in the DR52-associated DRB1 group: identification of DRB1*1115, DRB1*1117 and DRB1*1319.
Low and high resolution sequence specific oligonucleotide probe hybridization patterns were used to design an approach to direct sequencing of allele specific amplified cDNA. Several PCR amplifications were used to derive overlapping sequence fragments to define complete first domain sequences for a single allele. This method has been used to characterize three new DRB1 alleles in the DR52 family, DRB1*1115, DRB1*1117, and DRB1*1319. All three alleles carry polymorphisms previously observed in other DRB alleles and underscore the importance of utilizing a directed sequencing approach for obtaining unambiguous typing results in matching for bone marrow transplantation between unrelated donor and recipient.
Robbins
F
F
Department of Pediatrics, Georgetown University School of Medicine, Washington, DC, USA.
Tang
T
T
Yao
H
H
Ng
J
J
Hartzman
R J
RJ
Hurley
C K
CK
D000483:Alleles; D001483:Base Sequence; D002461:Cell Line, Transformed; D018076:DNA, Complementary; D014644:Genetic Variation; D006684:HLA-DR Antigens / Q000235:genetics*; D059811:HLA-DRB1 Chains; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012689:Sequence Homology, Nucleic Acid
D018076:DNA, Complementary; D006684:HLA-DR Antigens; D059811:HLA-DRB1 Chains