A homogeneous HLA-B*27 genotyping assay using dried reagent mixtures

Dis Markers. 2009;27(2):85-91. doi: 10.3233/DMA-2009-0653.

Abstract

The presence of HLA-B*27 allele with patients suspected with ankylosing spondylitis can be used in the diagnostic process. We have developed an assay for typing for the HLA-B*27 in whole blood dried on sample collection cards using pre-dried reagent wells and homogeneous time-resolved fluorescence based PCR approach. Essentially only the sample needs to be added to the dry ready-to-use reaction well in order to start the homogenous amplification assay. The method was validated with 229 samples also typed with an existing DELFIA-based method and results of both assays were 100% concordant. The dried reagents were shown to be stable at least up to eight weeks at room temperature without any decline in their performance.

MeSH terms

  • DNA / blood*
  • DNA / chemistry
  • DNA / genetics*
  • Genotype
  • HLA-B27 Antigen / genetics*
  • Humans
  • Polymerase Chain Reaction
  • Reagent Kits, Diagnostic
  • Spondylitis, Ankylosing / blood
  • Spondylitis, Ankylosing / diagnosis*
  • Spondylitis, Ankylosing / genetics*

Substances

  • HLA-B27 Antigen
  • Reagent Kits, Diagnostic
  • DNA