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Development of multiplex pyrosequencing for HLA-B*57:01 screening using single nucleotide polymorphism haplotype.

N Sankuntaw1, S Chantarangsu, W Chantratita

  • 1Department of Microbiology and Research, Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand.

Journal of Clinical Pharmacy and Therapeutics
|May 28, 2014
PubMed
Summary

A new multiplex pyrosequencing method accurately screens for HLA-B*57:01, a key genetic marker for abacavir hypersensitivity. This cost-effective and rapid test improves HIV-1 treatment safety for the Thai population.

Keywords:
adverse effectgenotypingpharmacogenetics

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Area of Science:

  • Genetics
  • Pharmacogenomics
  • Molecular Diagnostics

Background:

  • Abacavir (ABC) is a vital antiretroviral for HIV-1 treatment.
  • Mandatory HLA-B*57:01 screening precedes ABC initiation due to hypersensitivity risks.
  • Current screening methods are costly, slow, and require specialized labs.

Purpose of the Study:

  • To develop a more accurate, cost-effective, and rapid screening method for HLA-B*57:01.
  • To utilize multiplex pyrosequencing with rs2395029 and rs3093726 as surrogate markers.

Main Methods:

  • Developed multiplex pyrosequencing assay for HLA-B*57:01 screening.
  • Used rs2395029 and rs3093726 as surrogate markers.
  • Validated the assay in 130 Thai subjects against singleplex pyrosequencing and standard sequencing.

Main Results:

  • Multiplex pyrosequencing demonstrated 100% concordance with established methods.
  • Achieved 100% sensitivity, specificity, negative predictive value, and positive predictive value.
  • The assay proved accurate, cost-effective, and rapid.

Conclusions:

  • Multiplex pyrosequencing using rs2395029 and rs3093726 is a reliable method for HLA-B*57:01 screening.
  • This assay offers a powerful tool for abacavir hypersensitivity screening.
  • Applicable for pre-treatment screening in the Thai population, enhancing treatment safety.