HLA-B typing by reference strand mediated conformation analysis using a capillary-based semiautomated genetic analyzer

The application of reference strand conformation analysis (RSCA) to HLA-A t yping using the ABI PRISM 310 capillary based genetic analyzer has recently been described. This study outlines the development and validation of capi llary RSCA for HLA-B typing. Mobility values for 93 HLA-B alleles were defi ned following electrophoresis of known controls through the system. Three f luorescently labelled references, labelled with three different dyes can be electrophoresed simultaneously. The technique was validated by comparing r esults from 296 cord blood donors with those obtained using reverse SSO. Fo llowing capillary RSCA 14.5% of samples required confirmatory typing, compa red with a repeat rate of 5.1% following reverse SSO. In samples where no o ther typing was necessary there was 100% correlation between the two method s. Capillary RSCA for HLA-B typing is quick, easy to implement, and with th e introduction of new FLRs and gel matrices has the potential to evolve int o a high resolution typing method. (C) American Society for Histocompatibil ity and Immunogenetics, 2001. Published by Elsevier Science Inc.