HLA-A typing by reference strand-mediated conformation analysis (RSCA) using a capillary-based semi-automated genetic analyser.
HLA typing of class I loci by reference strand-mediated conformation analysis (RSCA) using a slab gel genetic analyser has been described. This study adapted the method for use in the capillary based ABI PRISM 310. Control DNA samples were used to create a database of mobility values for 37 HLA-A alleles. The technique was validated by comparing RSCA and sequence-specific oligonucleotide probe (SSOP)/sequence-specific primer amplification (SSP) HLA-A locus typing results from 214 cord blood samples. Of the samples tested, 6.5% required confirmatory typing by SSP, compared with a repeat rate of 10-40% for SSOP. In 200 samples where no SSP was necessary, there was 100% concordance between RSCA and previous results. The ABI PRISM 310 RSCA method defines HLA-A types at medium resolution and is quick and easy to implement.
Turner
D M
DM
Department of Histocompatibility and Immunogenetics, North London Centre, UK. david.turner@nbs.nhs.uk
Poles
A
A
Brown
J
J
Argüello
J R
JR
Madrigal
J A
JA
Navarrete
C V
CV
D019075:Electrophoresis, Capillary; D005069:Evaluation Studies as Topic; D005821:Genetic Techniques; D015234:HLA-A Antigens / Q000235:genetics*; D020180:Heteroduplex Analysis; D006650:Histocompatibility Testing / Q000379:methods* / Q000592:standards*; D006801:Humans; D017422:Sequence Analysis, DNA / Q000295:instrumentation* / Q000379:methods
D015234:HLA-A Antigens