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A Novel Multiplex HRM Assay to Detect Clopidogrel Resistance
Lichen Zhang1,2, Xiaowei Ma3, Guoling You2
1School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, 200127, P.R. China.
Scientific Reports
|November 24, 2017
Summary
A new multiplex assay accurately genotypes five key genetic markers linked to clopidogrel resistance. This method enhances personalized antiplatelet therapy for patients with cardiovascular conditions.
Area of Science:
- Pharmacogenomics
- Molecular Diagnostics
Background:
- Clopidogrel response varies due to genetic factors, impacting treatment efficacy.
- Five specific single nucleotide polymorphisms (SNPs) are linked to clopidogrel resistance in the Chinese population.
Purpose of the Study:
- To develop a multiplex high-resolution melting (HRM) assay for simultaneous genotyping of five clopidogrel resistance-associated SNPs.
- To evaluate the accuracy and efficiency of the developed multiplex HRM assay.
Main Methods:
- Design of multiplex amplicons with distinct melting temperatures using uMelt software and GC-rich tails.
- Validation of the multiplex HRM assay against Digital Fluorescence Molecular Hybridization (DFMH) and Sanger sequencing.
- Testing on 316 DFMH-pretested samples from acute coronary syndrome patients.
Main Results:
- The multiplex HRM assay successfully genotyped five SNPs in two tubes.
- Sensitivity was 100% and specificity was 99.93% for the HRM assay.
- The assay demonstrated high accuracy in detecting variants associated with clopidogrel resistance.
Conclusions:
- Multiplex HRM is a highly accurate, fast, and cost-effective closed-tube method for genotyping clopidogrel resistance SNPs.
- This assay can aid in optimizing clopidogrel therapy through personalized genetic screening.
- The developed method offers a valuable tool for clinical pharmacogenomic applications.
