HLA-DRB sequencing-based typing: an improved protocol which shows complete DRB exon 2 sequences and includes exon 3 of HLA-DRB4/5.
Several human leukocyte antigen (HLA)-DRB protocols for sequencing-based typing have been described. In general, the DRB1 amplification is performed using group-specific amplification primers (GSAPs) located in HVR I or intron 1. Only some protocols include amplification of DRB3, DRB4, and DRB5. However, prior knowledge obtained by alternative methods such as PCR-SSP is preferred for some protocols and a large amount of DNA is often required for adequate typing of HLA-DRB1. We describe a new protocol that uses GSAPs located in the introns, includes also sequencing of exon 3 to enable high resolution. This protocol allows DRB typing without prior knowledge of the HLA type.
van Dijk
A
A
Department of Pathology, University Medical Centre Utrecht, Utrecht, The Netherlands.
Melchers
R
R
Hilkes
Y
Y
Rozemuller
E
E
Tilanus
M
M
D017931:DNA Primers; D005091:Exons / Q000235:genetics*; D006684:HLA-DR Antigens / Q000235:genetics*; D059845:HLA-DRB4 Chains; D059847:HLA-DRB5 Chains; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D007438:Introns / Q000235:genetics; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D006684:HLA-DR Antigens; D059845:HLA-DRB4 Chains; D059847:HLA-DRB5 Chains