HLA-A typing by reference strand-mediated conformation analysis (RSCA) using a capillary-based semi-automated genetic analyser

Tissue Antigens. 1999 Oct;54(4):400-4. doi: 10.1034/j.1399-0039.1999.540410.x.

Abstract

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.

MeSH terms

  • Electrophoresis, Capillary
  • Evaluation Studies as Topic
  • Genetic Techniques
  • HLA-A Antigens / genetics*
  • Heteroduplex Analysis
  • Histocompatibility Testing / methods*
  • Histocompatibility Testing / standards*
  • Humans
  • Sequence Analysis, DNA / instrumentation*
  • Sequence Analysis, DNA / methods

Substances

  • HLA-A Antigens