Ligation based HLA-B*27 typing.
We have established a ligation based typing method to detect HLA-B*27 alleles at the DNA level. The method requires amplification of exon 2 of the HLA-B locus from genomic DNA by the polymerase catalyzed chain reaction (PCR) using group specific primers. An aliquot of the PCR amplification product, heat stable ligase and a pair of oligonucleotide probes, designed to hybridize adjacently to HLA-B*27 specific sequences of the amplified DNA are subsequently thermocycled. If the probes are perfectly complementary they become ligated otherwise they stay separated. The ligation of probes can be detected through their different labels by an enzyme linked immunosorbent assay (ELISA). Ligation based detection of beta-actin sequences which have been co-amplified serves as positive control for each PCR reaction. We observed complete concordance when typing 76 HLA-B*27 positive and 107 HLA-B*27 negative individuals either by serology or by the ligation based approach. We conclude that ligation based typing is a reliable tool for the DNA based detection of HLA-B*27 alleles. The procedure allows automation to a large extent and should be easily applicable to the typing of other HLA-class I alleles.
Department of Blood Group Serology, University of Vienna.
G F
GF
Fischer
I
I
Faé
S
S
Moser
M
M
Petrasek
H M
HM
Bässler
G R
GR
Menzel
W R
WR
Mayr
D001483:Base Sequence / Q000235:genetics; D005838:Genotype; D015796:HLA-B27 Antigen / Q000235:genetics*; D006801:Humans; D007963:Leukocytes, Mononuclear; D008967:Molecular Biology / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods
D015796:HLA-B27 Antigen