HLA-B*5701 typing by sequence-specific amplification: validation and comparison with sequence-based typing.
Susceptibility to abacavir hypersensitivity (ABC HSR) is strongly associated with alleles carried on the 57.1 ancestral haplotype including HLA-B*5701 and Hsp70 Hom M493T. In one study, prospective testing for HLA-B*5701 and exclusion of individuals carrying this allele, from receiving abacavir, substantially lowered the incidence of ABC HSR to 0% (95% confidence interval 0-0.075%). The presence of HLA-B*5701 is usually detected by standard serological tests and by molecular genetic methods such as sequence-based typing (SBT). While the former test cannot discriminate between HLA-B57 subtypes, the expensive SBT may not be readily available in all laboratories. Hence, an alternate method was developed to detect HLA-B*5701 using allele and group-specific polymerase chain reaction-sequence-specific primers (PCR-SSP) typing. This PCR-SSP-typing method positively amplified all HLA-B*5701 alleles in concordance with their SBT-assigned typing. This multiplexed SSP assay was able to distinguish between HLA-B*5701 (n = 10) and closely related HLA-B57 alleles B*5702 (n = 2), -B*5703 (n = 1), -B*5704 (n = 1) alleles and non-HLA-B*57 alleles (n = 61). In conclusion, this method of HLA-B*5701 detection is a rapid and accurate typing method with high specificity, sensitivity and reproducibility.
Martin
A M
AM
Center for Clinical Immunology and Biomedical Genetics, Royal Perth Hospital and Murdoch University, Western Australia 6000. annalise.martin@health.wa.gov.au
Nolan
D
D
Mallal
S
S
D000483:Alleles; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D015224:Dideoxynucleosides / Q000494:pharmacology; D020022:Genetic Predisposition to Disease; D015235:HLA-B Antigens / Q000096:biosynthesis* / Q000235:genetics*; D006239:Haplotypes; D006650:Histocompatibility Testing / Q000379:methods*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D018894:Reverse Transcriptase Inhibitors / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D000099227:Abacavir
D017931:DNA Primers; D015224:Dideoxynucleosides; D015235:HLA-B Antigens; D018894:Reverse Transcriptase Inhibitors; D000099227:Abacavir; C484830:HLA-B*57:01 antigen