A novel multiplex polymerase chain reaction assay for detection of both HLA-A*31:01/HLA-B*15:02 alleles, which confer susceptibility to carbamazepine-induced severe cutaneous adverse reactions

HLA. 2017 Dec;90(6):335-342. doi: 10.1111/tan.13143. Epub 2017 Oct 2.

Abstract

HLA-A*31:01 and HLA-B*15:02 have been widely reported to confer genetic susceptibility to carbamazepine (CBZ)-induced severe cutaneous adverse reactions (SCARs). Accordingly, the screening for these alleles has been highly recommended to prevent SCAR prior to introducing CBZ therapy. Although a number of methods are available for screening of HLA-A*31:01 or HLA-B*15:02 alleles separately, developing an assay that can detect both these alleles would be more clinically practical, cost-effective and less time-consuming. Therefore, in this study, a multiplex polymerase chain reaction (PCR) using TaqMan Probe was designed and validated to be able to detect HLA-A*31:01 and HLA-B*15:02. In comparison with Luminex-SSO/SBT/SSB, the multiplex PCR assay for detection of HLA-A*31:01 and HLA-B*15:02 had a perfect agreement in the validation group of 125 samples. The method was able to detect the target genes at the DNA concentration of 0.037 ng/μL. The unit cost of this assay is less than $5 USD with total time of 110 minutes.

Keywords: Stevens-Johnson Syndrome; TaqMan Probe; carbamazepine; drug rash with eosinophilia and systemic symptoms; real-time PCR; severe cutaneous adverse reactions.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Anticonvulsants / adverse effects*
  • Base Sequence
  • Carbamazepine / adverse effects*
  • Drug Hypersensitivity Syndrome / diagnosis
  • Drug Hypersensitivity Syndrome / etiology
  • Drug Hypersensitivity Syndrome / genetics*
  • Drug Hypersensitivity Syndrome / immunology
  • Gene Expression
  • Genetic Predisposition to Disease
  • HLA-A Antigens / genetics*
  • HLA-A Antigens / immunology
  • HLA-B15 Antigen / genetics*
  • HLA-B15 Antigen / immunology
  • Humans
  • Limit of Detection
  • Multiplex Polymerase Chain Reaction / economics
  • Multiplex Polymerase Chain Reaction / methods*
  • Reproducibility of Results
  • Sequence Alignment
  • Skin / drug effects
  • Skin / immunology
  • Skin / pathology
  • Stevens-Johnson Syndrome / diagnosis
  • Stevens-Johnson Syndrome / etiology
  • Stevens-Johnson Syndrome / genetics*
  • Stevens-Johnson Syndrome / immunology

Substances

  • Anticonvulsants
  • HLA-A Antigens
  • HLA-A*31:01 antigen
  • HLA-B*15:02 antigen
  • HLA-B15 Antigen
  • Carbamazepine