Allele-specific HLA-DR typing by mass spectrometry: an alternative to hybridization-based typing methods.
The primer oligomer base extension (PROBE) reaction, combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, is used to characterize HLA-DR2 polymorphism. Alleles are distinguished rapidly and accurately by measuring the mass of primer extension products at every known variable region of HLA-DR2 alleles. Since differentiation of alleles by PROBE relies on measuring differences in extension product mass rather than differences in hybridization properties, mistyped alleles resulting from nonspecific hybridization are absent. The method shows considerable potential for high-throughput screening of HLA-DR polymorphism in a chip-based format, including rapid tissue typing of unrelated volunteer donors.
Worrall
T A
TA
Department of Pharmacology, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
Schmeckpeper
B J
BJ
Corvera
J S
JS
Cotter
R J
RJ
D000483:Alleles; D003062:Codon; D004247:DNA / Q000097:blood; D006684:HLA-DR Antigens / Q000032:analysis* / Q000235:genetics; D006650:Histocompatibility Testing / Q000379:methods*; D006801:Humans; D017403:In Situ Hybridization; D016133:Polymerase Chain Reaction; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003062:Codon; D006684:HLA-DR Antigens; D004247:DNA