Vitamin K Epoxide Reductase Complex (VKORC1) Electrochemical Genosensors: Towards the Identification of 1639 G>A Genetic Polymorphism

Anticoagulants, including warfarin, are often administered to patients who are exhibiting early symptoms of thromboembolic episodes or who have already experienced such episodes. However, warfarin has a limited therapeutic index and might cause bleeding and other clinical problems. Warfarin inhibits the vitamin K epoxide reductase complex subunit 1 (VKORC1), an enzyme essential for activating vitamin K, in the coagulation cascade. Genetic factors, such as polymorphisms, can change the natural function of VKORC1, causing variations in the medication reaction among individuals. Hence, before prescribing warfarin, the patient’s genetic profile should also be considered. In this study, an electrochemical genosensor capable of detecting the VKORC1 1639 G>A polymorphism was designed and optimized. This analytical approach detects the electric current obtained during the hybridization reaction between two 52 base pair complementary oligonucleotide sequences. Investigating public bioinformatic platforms, two DNA sequences with the A and G single-nucleotide variants were selected and designed. The experimental protocol of the genosensor implied the formation of a bilayer composed of a thiolate DNA and an alkanethiol immobilized onto gold electrodes, as well as the formation of a DNA duplex using a sandwich-format hybridization reaction through a fluorescein labelled DNA signalling probe and the enzymatic amplification of the electrochemical signal, detected by chronoamperometry. A detection limit of 20 pM and a linear range of 0.05–1.00 nM was obtained. A clear differentiation between A/A, G/A and G/G genotypes in biological samples was successfully identified by his novel device.

Keywords

Cardiovascular diseases Electrochemical genosensors Single-nucleotide polymorphisms VKORC1 Warfarin

URI

http://hdl.handle.net/10400.22/30220

Citation

Barbosa, T., Morais, S. L., Carvalho, R., Magalhães, J. M. C. S., Domingues, V. F., Delerue-Matos, C., Ferreira-Fernandes, H., Pinto, G. R., Santos, M., & Barroso, M. F. (2025). Vitamin K Epoxide Reductase Complex (VKORC1) Electrochemical Genosensors: Towards the Identification of 1639 G>A Genetic Polymorphism. Chemosensors, 13(7), Artigo 7. https://doi.org/10.3390/chemosensors13070248