Rapid, Reliable, and Inexpensive HLA-B*58:01 Detection Method Using DNA Binding Dye-based Duplex Allele-specific Melting Curve Analysis

Ann Clin Lab Sci. 2018 May;48(3):296-300.

Abstract

Background: Allopurinol is the most commonly used drug for the treatment of gout and also one of the most common causes of severe cutaneous adverse reactions (SCARs). Human leukocyte antigen-B*58:01 (HLA-B*58:01) is strongly associated with allopurinol-induced SCARs. The aim of the present study was to develop and validate a rapid and economic screening method for HLA-B*58:01.

Methods: The accuracy of duplex allele-specific melting curve analysis using DNA-binding dye for HLA-B*58:01 was evaluated in 150 blood samples with sequence-based typing (SBT) as the reference method.

Results: Fifty HLA-B*58:01-positive and 100 negative results obtained by duplex allele-specific melting curve analysis were completely in agreement with the SBT results.

Conclusion: Duplex allele-specific melting curve analysis is a rapid, reliable and inexpensive assay that is appropriate for screening for the HLA-B*58:01 allele.

Keywords: Allopurinol hypersensitivity; DNA-binding dye; HLA-B*58:01; Melting curve analysis; Severe cutaneous adverse reactions.

MeSH terms

  • Alleles
  • DNA / genetics
  • DNA / metabolism*
  • Genotyping Techniques / methods*
  • HLA-B Antigens / analysis*
  • HLA-B Antigens / genetics*
  • Humans
  • Predictive Value of Tests
  • Real-Time Polymerase Chain Reaction / methods*
  • Transition Temperature*

Substances

  • HLA-B Antigens
  • DNA