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HLA-B*5701 typing by sequence-specific amplification: validation and comparison with sequence-based typing.

A M Martin·D Nolan·S Mallal
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Abstract

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.

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AllelesBase SequenceDNA PrimersDideoxynucleosidesGenetic Predisposition to DiseaseHLA-B AntigensHaplotypesHistocompatibility TestingMolecular Sequence DataPolymerase Chain ReactionPolymorphism, Single-Stranded ConformationalReverse Transcriptase InhibitorsSequence Analysis, DNAAbacavirJournal ArticleResearch Support, Non-U.S. Gov't

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A M Martin, D Nolan, S Mallal. 2005. HLA-B*5701 typing by sequence-specific amplification: validation and comparison with sequence-based typing.. https://doi.org/10.1111/j.1399-0039.2005.00401.x

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