Validation of a Rapid, Robust, Inexpensive Screening Method for Detecting the HLA-B*58:01 Allele in the Prevention of Allopurinol-Induced Severe Cutaneous Adverse Reactions

Dinh Van Nguyen; Christopher Vida; Hieu Chi Chu; Richard Fulton; Jamma Li; Suran L Fernando

doi:10.4168/aair.2017.9.1.79

Validation of a Rapid, Robust, Inexpensive Screening Method for Detecting the HLA-B*58:01 Allele in the Prevention of Allopurinol-Induced Severe Cutaneous Adverse Reactions

Allergy Asthma Immunol Res. 2017 Jan;9(1):79-84. doi: 10.4168/aair.2017.9.1.79.

Authors

Dinh Van Nguyen^{1

2

3}, Christopher Vida⁴, Hieu Chi Chu⁵, Richard Fulton², Jamma Li^{2

6}, Suran L Fernando^{1

2

6}

Affiliations

¹ Sydney Medical School - Northern, University of Sydney, Sydney, Australia.
² ImmunoRheumatology Laboratory, Pathology North-Northern Sydney, St Leonards, Australia.
³ Department of Allergy and Clinical Immunology, Hanoi Medical University, Hanoi, Vietnam. vngu3162@uni.sydney.edu.au.
⁴ ImmunoRheumatology Laboratory, Pathology North-Northern Sydney, St Leonards, Australia. christophervid@gmail.com.
⁵ Center of Allergology and Clinical Immunology, Bach Mai Hospital, Hanoi, Vietnam.
⁶ Department of Clinical Immunology and Allergy, Royal North Shore Hospital, Sydney, Australia.

Abstract

The HLA B*58:01 allele has been worldwide reported as a pharmacogenetic susceptibility to allopurinol-induced severe cutaneous adverse reactions (SCARs). To prevent these life-threatening conditions, the American College of Rheumatology hingly recommended that the HLA-B*58:01 be screened prior to the initiation of allopurinol therapy. Therefore, we developed a rapid, robust, inexpensive screening method using SYBR® Green real time PCR to detect the HLA-B*58:01 allele. A total of 119 samples were tested. The assay has a sensitivity of 100% (95% CI: 69.15%-100%), a specificity of 100% (95% CI: 96.67%-100%), a positive predictive value of 100% (95% CI: 69.15%-100%) and a negative predictive value of 100% (95% CI: 96.67%-100%). HLA-B*58:01 genotyping results showed 100% agreement with those obtained from Luminex SSO/SBT/SSP. The lowest limit of detection of this method is 0.8 ng/μL of DNA. The unit cost of the test is only $3.8 USD. This novel screening test using SYBR® real time PCR would be appropriate to identify individuals with the HLA-B*58:01 allele for the prevention of allopurinol-induced SCARs.

Keywords: Allopurinol hypersensitivity; HLA-B antigens; Stevens Johnson syndrome; drug reactions with eosinophilia and systemic symptoms; real time polymerase chain reaction; severe cutaneous adverse reactions; toxic epidermal necrolysis.