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Candidate Gene Testing in Clinical Cohort Studies with Multiplexed Genotyping and Mass Spectrometry
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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
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.
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.
