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

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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.

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