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Genetic variations significantly influence drug response through pharmacokinetics, receptor interactions, and biologic milieu modifications. Pharmacokinetic alterations impact drug metabolism and clearance, affecting efficacy and toxicity. Variants in drug-metabolizing enzymes, such as CYP2C9 and CYP2C19, alter drug activation and elimination. For example, CYP2C9 loss-of-function variants require lower warfarin doses to prevent excessive bleeding, while CYP2C19 variants reduce clopidogrel...
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    Area of Science:

    • Pharmacogenomics
    • Molecular Diagnostics
    • Biosensor Technology

    Background:

    • CYP2C19 loss-of-function alleles (*2, *3, *4, *5) impair clopidogrel activation, leading to reduced efficacy and adverse cardiovascular outcomes.
    • Genetic testing for these polymorphisms is essential for personalized antiplatelet therapy.

    Purpose of the Study:

    • To develop a novel biosensor-based microarray for the visual detection of CYP2C19 loss-of-function polymorphisms.
    • To establish a rapid, accurate, and accessible method for clinical genetic testing.

    Main Methods:

    • DNA amplification using 5'-biotinylated primers, followed by reversed hybridization on a gene chip.
    • Detection via horseradish peroxidase-conjugated streptavidin and colorimetric development.
    • Validation using constructed plasmids and comparison with direct sequencing in 88 clinical samples.

    Main Results:

    • The assay provides visual results detectable by the naked eye.
    • Perfect concordance (κ=1.000) with plasmids for alleles *2, *3, *4, and *5.
    • High concordance (κ=0.983) with direct sequencing in clinical samples, demonstrating excellent accuracy and a low detection limit (5x10² cells/mL).

    Conclusions:

    • The developed biosensor microarray enables in situ signal amplification for easy detection.
    • This method is highly accurate, specific, and suitable for routine clinical application in hospital laboratories.
    • It offers a promising tool for guiding clopidogrel therapy based on individual CYP2C19 genotype.