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1. Electrophoresis. 2012 Apr;33(8):1306-10. doi: 10.1002/elps.201100457.

Rapid testing of clopidogrel resistance by genotyping of CYP2C19 and CYP2C9
polymorphisms using denaturing on-chip capillary electrophoresis.

Minarik M, Kopeckova M, Gassman M, Osmancik P, Benesova L.

Center for Applied Genomics of Solid Tumors, Genomac Research Institute, Prague,
Czech Republic.

Antiplatelet therapy is a cornerstone of cardiovascular treatment in patients
with coronary artery disease and after myocardial infarction. Clopidogrel has
become a popular antiplatelet agent due to its fast action and low frequency of
adverse effects. Kinetics of clopidogrel metabolism is driven by enzymatic
activity of the Cytochrome P450 system. Genotyping of CYP2C19 and CYP2C9
polymorphisms allows to identify slow metabolizers showing resistance to
clopidogrel therapy. Today, a number of PCR-based techniques for single
nucleotide polymorphism genotyping directed at clopidogrel resistance
polymorphisms are in use. Here, we describe a new alternative genotyping approach
combining the separation power of denaturing capillary electrophoresis with the
analysis speed and ease of use of Bioanalyzer chipCE platform. Using an upgraded
heater control, we present an optimization for allele separation of CYP2C19
I331V, CYP2C9 R144C, and CYP2C9 I359L polymorphisms employing run temperatures of
up to 55°C. We demonstrate rapid and accessible approach to reproducible
clopidogrel resistance with feasibility and low cost.

© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

PMID: 22589111 [PubMed - in process]