title: Detection of HLA-B*58:01 with TaqMan assay and its association with allopurinol-induced sCADR.
abstract: BACKGROUND
  The HLA-B*58:01 allele is associated with allopurinol-induced severe cutaneous adverse drug reactions (sCADR) in certain geographic regions, but the diversity of the correlation is large. In addition, the currently available HLA-B*58:01 testing methods are too laborious for use in routine clinical detection. The objective of this study was to develop a new, convenient method for the detection of HLA-B*58:01 and to investigate the association of HLA-B*58:01 with allopurinol-induced sCADR in a Han Chinese population.
  
  
  METHODS
  A new method combining sequence-specific primers (SSP) and TaqMan probe amplification was developed in this study and was used to detect the HLA-B*58:01 in 48 allopurinol-induced sCADR, 133 allopurinol-tolerant, and 280 healthy individuals. The accuracy, sensitivity, and specificity were assessed by a commercial PCR-SSP HLA-B typing kit. The low limit of detection was detected by serial dilution of an HLA-B*58:01-positive DNA template.
  
  
  RESULTS
  The new method successfully identified HLA-B*58:01 in thousands of HLA-B alleles, and the results for 344 DNA samples were perfectly concordant with the results of the commercial PCR-SSP HLA-B kit. The analytical sensitivity is 100% and the specificity is over 99%. The low limit of detection of this assay is 100 pg DNA, which was 10 times more sensitive than the commercial PCR-SSP kit. HLA-B*58:01 was present in 93.8% of the patients with sCADR, 7.5% of the allopurinol-tolerant patients, and 12.1% of the healthy controls. The frequency of HLA-B*58:01 was significantly higher in the sCADR group than in the control group (p<0.0001). However, there was no significant difference between the allopurinol-tolerant and control groups (p=0.1547).
  
  
  CONCLUSIONS
  HLA-B*58:01 has a strong association with allopurinol-induced sCADR in Han Chinese. The newly developed method is reliable for HLA-B*58:01 detection prior to allopurinol therapy.
authors.forename: Xinju
authors.initials: X
authors.lastname: Zhang
authors.forename: Huili
authors.initials: H
authors.lastname: Ma
authors.forename: Chunying
authors.initials: C
authors.lastname: Hu
authors.forename: Bo
authors.initials: B
authors.lastname: Yu
authors.forename: Weizhe
authors.initials: W
authors.lastname: Ma
authors.forename: Zhiyuan
authors.initials: Z
authors.lastname: Wu
authors.forename: Xiaoqun
authors.initials: X
authors.lastname: Luo
authors.forename: Hejian
authors.initials: H
authors.lastname: Zou
authors.forename: Ming
authors.initials: M
authors.lastname: Guan
mesh_terms: D000493:Allopurinol / Q000009:adverse effects*; D002681:China; D064420:Drug-Related Side Effects and Adverse Reactions / Q000235:genetics*; D015235:HLA-B Antigens / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012871:Skin Diseases / Q000139:chemically induced* / Q000235:genetics*
chemical_list: D015235:HLA-B Antigens; C500126:HLA-B*58:01 antigen; D000493:Allopurinol