title: Validation of a novel real-time PCR assay for detection of HLA-B*15:02 allele for prevention of carbamazepine - Induced Stevens-Johnson syndrome/Toxic Epidermal Necrolysis in individuals of Asian ancestry.
abstract: OBJECTIVE
  Screening for the HLA-B*15:02 allele has been recommended to prevent carbamazepine (CBZ) - induced Stevens-Johnson syndrome (SJS) and Toxic Epidermal Necrolysis (TEN) in individuals with Asian ancestry. We aimed, therefore, to develop and validate a robust and inexpensive method for detection of the HLA-B*15:02 allele.
  
  
  METHODS
  Real-time PCR using TaqMan® probes followed by SYBR® Green was used to detect the HLA-B*15:02 allele prior to treatment with CBZ therapy.
  
  
  RESULTS
  A total of 121 samples were tested. The assay has a sensitivity of 100% (95% CI: 76.84-100.0%), a specificity of 100% (95% CI: 96.61-100%), a positive predictive value of 100% (95% CI: 76.84-100%) and a negative predictive value of 100.0% (95% CI: 96.61-100.0%), respectively. There was 100% agreement between our results and genotyping using Luminex SSO/SBT/SSP. The lowest limit of detection of the TaqMan® probe is 0.05ng/μl and the SYBR® Green is 0.5ng/μl of DNA. The unit cost of using the TaqMan® probe followed by SYBR® Green is only $4.7 USD.
  
  
  CONCLUSIONS
  We developed a novel assay for the detection of the HLA-B*15:02 allele, which is robust, inexpensive and suitable for screening individuals of Asian ancestry in the prevention of CBZ-induced SJS/TEN.
authors.affiliation: ImmunoRheumatology Laboratory, Pathology North-Sydney, St Leonards, Australia; Sydney Medical School - Northern, University of Sydney, Sydney, Australia; Department of Allergy and Clinical Immunology, Hanoi Medical University, Hanoi, Viet Nam. Electronic address: vngu3162@uni.sydney.edu.au.
authors.forename: Dinh Van
authors.initials: DV
authors.lastname: Nguyen
authors.affiliation: ImmunoRheumatology Laboratory, Pathology North-Sydney, St Leonards, Australia. Electronic address: christophervid@gmail.com.
authors.forename: Christopher
authors.initials: C
authors.lastname: Vidal
authors.affiliation: Center of Allergology and Clinical Immunology, Bach Mai Hospital, Hanoi, Viet Nam.
authors.forename: Hieu Chi
authors.initials: HC
authors.lastname: Chu
authors.affiliation: Department of Immunology and Molecular Biology, National Institute of Hygiene and Epidemiology, Hanoi, Viet Nam.
authors.forename: Nga Thi Quynh
authors.initials: NT
authors.lastname: Do
authors.affiliation: Hanoi Heart Hospital, Hanoi, Viet Nam.
authors.forename: Tu Thi Linh
authors.initials: TT
authors.lastname: Tran
authors.affiliation: Department of Allergy, Immunology and Rheumatology, National Hospital of Pediatrics, Hanoi, Viet Nam.
authors.forename: Huong Thi Minh
authors.initials: HT
authors.lastname: Le
authors.affiliation: ImmunoRheumatology Laboratory, Pathology North-Sydney, St Leonards, Australia.
authors.forename: Richard B
authors.initials: RB
authors.lastname: Fulton
authors.affiliation: ImmunoRheumatology Laboratory, Pathology North-Sydney, St Leonards, Australia; Department of Clinical Immunology and Allergy, Royal North Shore Hospital, Sydney, Australia.
authors.forename: Jamma
authors.initials: J
authors.lastname: Li
authors.affiliation: ImmunoRheumatology Laboratory, Pathology North-Sydney, St Leonards, Australia; Sydney Medical School - Northern, University of Sydney, Sydney, Australia; Department of Clinical Immunology and Allergy, Royal North Shore Hospital, Sydney, Australia.
authors.forename: Suran L
authors.initials: SL
authors.lastname: Fernando
mesh_terms: D000483:Alleles; D044466:Asian People; D002220:Carbamazepine / Q000009:adverse effects* / Q000627:therapeutic use; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing; D005838:Genotype; D059910:HLA-B15 Antigen / Q000235:genetics*; D006801:Humans; D011110:Polymorphism, Genetic; D011237:Predictive Value of Tests; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012016:Reference Values; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013262:Stevens-Johnson Syndrome / Q000175:diagnosis* / Q000517:prevention & control
chemical_list: C483536:HLA-B*15:02 antigen; D059910:HLA-B15 Antigen; D002220:Carbamazepine
keywords: Carbamazepine hypersensitivity; HLA-B antigens; Real-time polymerase chain reaction; SCARs; Severe cutaneous adverse reactions; Stevens Johnson syndrome; Toxic Epidermal Necrolysis