{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,13]],"date-time":"2026-01-13T21:53:28Z","timestamp":1768341208619,"version":"3.49.0"},"reference-count":18,"publisher":"MDPI AG","issue":"4","license":[{"start":{"date-parts":[[2025,3,24]],"date-time":"2025-03-24T00:00:00Z","timestamp":1742774400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"CYTED-rede Ibero-Americana project","award":["GENOPSYSEN-ref: 223RT0141"],"award-info":[{"award-number":["GENOPSYSEN-ref: 223RT0141"]}]},{"name":"FCT\/MECI, Funda\u00e7\u00e3o para a Ci\u00eancia e Tecnologia and Minist\u00e9rio da Educa\u00e7\u00e3o, Ci\u00eancia e Inova\u00e7\u00e3o","award":["GENOPSYSEN-ref: 223RT0141"],"award-info":[{"award-number":["GENOPSYSEN-ref: 223RT0141"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Background\/Objectives: The CYP2C9 enzyme is involved in the metabolism of warfarin. The CYP2C9 gene harbors several single-nucleotide polymorphisms (SNPs), including CYP2C9*2 (rs1799853), which is known to affect warfarin\u2019s therapeutic response. So, it is important to develop analytical tools capable of genotyping these SNPs to adjust warfarin\u2019s therapeutic outcomes. In this work, an electrochemical DNA-based sensor was constructed and optimized for the detection of the CYP2C9*2 polymorphism. Methods: Using bioinformatic database platforms, two 71 base pair DNA target probes with the polymorphic variants A and G were chosen and designed. A DNA-based sensor was composed by mercaptohexanol and the CYP2C9*2 DNA capture probe in a self-assembled monolayer connected to screen-printed gold electrodes. Two independent hybridization events of the CYP2C9*2 allele were designed using complementary fluorescein-labeled DNA signaling to improve selectivity and avoid secondary structures. Three human samples with the homozygous variant (G\/G) and non-variant (A\/A) and heterozygous (G\/A) genotypes were amplified by PCR and then applied to the developed genosensor. Results: Chronoamperometry measurements were performed for both polymorphic probes. A calibration curve in the 0.25 to 2.50 nM (LOD of 13 pM) and another in the 0.15 to 5.00 nM range (LOD of 22.6 pM) were obtained for the homozygous non-variant and variant probes, respectively. This innovative tool was capable of identifying the hybridization reaction between two complementary strands of immobilized DNA, representing a genotyping alternative to the classical PCR methodology. Conclusions: The developed electrochemical DNA-based sensor was able to discriminate two synthetic SNP target sequences (Target-A and Target-G) and detect, with specificity, the three patients\u2019 genotypes (G\/G, G\/A, and A\/A). This tool is therefore a promising, sensitive, and cost-effective analytical way to determine and discriminate an individual\u2019s genotype and predict the appropriate warfarin dose.<\/jats:p>","DOI":"10.3390\/genes16040372","type":"journal-article","created":{"date-parts":[[2025,3,24]],"date-time":"2025-03-24T13:48:20Z","timestamp":1742824100000},"page":"372","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Warfarin Pharmacogenomics: Designing Electrochemical DNA-Based Sensors to Detect CYP2C9*2 Gene Variation"],"prefix":"10.3390","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-7654-3095","authenticated-orcid":false,"given":"Tiago","family":"Barbosa","sequence":"first","affiliation":[{"name":"REQUIMTE\/LAQV, Escola Superior de Sa\u00fade, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 400, 4200-072 Porto, Portugal"},{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 431, 4249-015 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0341-4511","authenticated-orcid":false,"given":"Stephanie L.","family":"Morais","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 431, 4249-015 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2721-7069","authenticated-orcid":false,"given":"Eduarda","family":"Pereira","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Escola Superior de Sa\u00fade, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 400, 4200-072 Porto, Portugal"},{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 431, 4249-015 Porto, Portugal"}]},{"given":"J\u00falia M. C. S.","family":"Magalh\u00e3es","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Departamento de Engenharia Qu\u00edmica, Faculdade de Engenharia, Universidade do Porto, Rua Dr. Roberto Frias, 4200-465 Porto, Portugal"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3472-849X","authenticated-orcid":false,"given":"Valentina F.","family":"Domingues","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 431, 4249-015 Porto, Portugal"}]},{"given":"Hygor","family":"Ferreira-Fernandes","sequence":"additional","affiliation":[{"name":"Instituto de Educa\u00e7\u00e3o, Ci\u00eancia e Tecnologia Do Piau\u00ed (IFPI), Departamento de Informa\u00e7\u00e3o, Ambiente, Sa\u00fade e Produ\u00e7\u00e3o Aliment\u00edcia, 64019-368 Teresina, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9110-1840","authenticated-orcid":false,"given":"Giovanny","family":"Pinto","sequence":"additional","affiliation":[{"name":"Grupo de Estudos Em Gen\u00e9tica Humana e M\u00e9dica (GEHMED), Laborat\u00f3rio de Gen\u00e9tica e Biologia Molecular, Departamento de Biomedicina, Universidade do Delta do Parna\u00edba (UFDPar), 64202-020 Parna\u00edba, Brazil"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5020-5942","authenticated-orcid":false,"given":"Marlene","family":"Santos","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Escola Superior de Sa\u00fade, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 400, 4200-072 Porto, Portugal"},{"name":"Grupo de Oncologia Molecular e Patologia Viral, Centro de Investiga\u00e7\u00e3o, Instituto Portugu\u00eas de Oncologia do Porto\u2014Francisco Gentil, Rua Dr. Ant\u00f3nio Bernardino de Almeida 865, 4200-072 Porto, Portugal"}]},{"given":"Maria F\u00e1tima","family":"Barroso","sequence":"additional","affiliation":[{"name":"REQUIMTE\/LAQV, Instituto Superior de Engenharia do Porto, Instituto Polit\u00e9cnico do Porto, Rua Dr. Ant\u00f3nio Bernardino de Almeida, 431, 4249-015 Porto, Portugal"}]}],"member":"1968","published-online":{"date-parts":[[2025,3,24]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"175","DOI":"10.1042\/ETLS20190197","article-title":"The future of precision medicine: Towards a more predictive personalized medicine","volume":"4","author":"Elemento","year":"2020","journal-title":"Emerg. Top. Life Sci."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"952","DOI":"10.1016\/j.fertnstert.2018.05.006","article-title":"Personalized medicine: Motivation, challenges and progress","volume":"109","author":"Goetz","year":"2018","journal-title":"Fertil. Steril."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1111\/cts.12884","article-title":"Precision Medicine, AI and the Future of Personalized Health Care","volume":"14","author":"Johnson","year":"2021","journal-title":"Clin. Transl. Sci."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1016\/j.cll.2016.05.006","article-title":"Fundamentals of Pharmacogenetics in Personalized, Precision Medicine","volume":"36","author":"Valdes","year":"2016","journal-title":"Clin. Lab. Med."},{"key":"ref_5","first-page":"13","article-title":"The role of pharmacogenetics and pharmacogenomics in improving translational medicine","volume":"6","author":"Dere","year":"2009","journal-title":"Clin. Cases Miner. Bone Metab."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"124692","DOI":"10.1016\/j.talanta.2023.124692","article-title":"Development of an electrochemical DNA-based biosensor for the detection of the cardiovascular pharmacogenetic-altering SNP CYP2C9*3","volume":"264","author":"Morais","year":"2023","journal-title":"Talanta"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Zhang, Q., Qi, Y., Wang, S., Zhao, F., Zou, L., Zhou, Q., Geng, P., Hong, Y., Yang, H., and Luo, Q. (2023). Identification and in vitro functional assessment of 10 CYP2C9 variants found in Chinese Han subjects. Front. Endocrinol., 14.","DOI":"10.3389\/fendo.2023.1139805"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1038\/s41397-020-0157-2","article-title":"CYP2C9 and VKORC1 genotyping for the quality of long-standing warfarin treatment in Russian patients","volume":"20","author":"Panchenko","year":"2020","journal-title":"Pharmacogenomics J."},{"key":"ref_9","unstructured":"Morais, S.L., Magalh\u00e3es, J., Domingues, V., Delerue-Matos, C., Ramos-Jesus, J., Ferreira-Fernandes, H., Pinto, G.R., and Barroso, M.F. (2021, January 1\u201315). Development of electrochemical genosensors applied to cardiovasular pharmacogenetics. Proceedings of the 1st International Electronic Conference on Chemical Sensors and Analytical Chemistry, Virtual."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"465","DOI":"10.2217\/pgs.12.2","article-title":"Clinical utility of pharmacogenetic biomarkers in cardiovascular therapeutics: A challenge for clinical implementation","volume":"13","author":"Ong","year":"2012","journal-title":"Pharmacogenomics"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Karki, R., Pandya, D., Elston, R.C., and Ferlini, C. (2015). Defining \u201cmutation\u201d and \u201cpolymorphism\u201d in the era of personal genomics. BMC Med. Genom., 8.","DOI":"10.1186\/s12920-015-0115-z"},{"key":"ref_12","unstructured":"Pratt, V.M., Scott, S.A., Pirmohamed, M., Esquivel, B., Kattman, B.L., and Malheiro, A.J. (2024, December 01). Warfarin Therapy and VKORC1 and CYP Genotype, Medical Genetics Summaries, Available online: https:\/\/www.ncbi.nlm.nih.gov\/books\/NBK84174\/."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1642","DOI":"10.1016\/j.bios.2006.07.015","article-title":"DNA single-base mismatch study with an electrochemical enzymatic genosensor","volume":"22","year":"2007","journal-title":"Biosens. Bioelectron."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"377","DOI":"10.4103\/1735-5362.235165","article-title":"The effect of CYP2C9*2, CYP2C9*3 and VKORC1-1639 G>A polymorphism in patients under warfarin therapy in city of Kermanshah","volume":"13","author":"Hosseinkhani","year":"2018","journal-title":"Res. Pharm. Sci."},{"key":"ref_15","first-page":"152","article-title":"Polymerase Chain Reaction and Its Application in the Diagnosis of Infectious Keratitis","volume":"8","author":"Liu","year":"2019","journal-title":"Med. Hypothesis Discov. Innov. Ophthalmol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"126877","DOI":"10.1016\/j.snb.2019.126877","article-title":"Allele-specific ligation and recombinase polymerase amplification for the detection of single nucleotide polymorphisms","volume":"298","author":"Yamanaka","year":"2019","journal-title":"Sens. Actuators B Chem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"249","DOI":"10.1007\/s13246-016-0519-x","article-title":"DNA sensors to assess the effect of VKORC1 and CYP2C9 gene polymorphisms on warfarin dose requirement in Chinese patients with atrial fibrillation","volume":"40","author":"Huang","year":"2017","journal-title":"Australas. Phys. Eng. Sci. Med."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"e13","DOI":"10.1016\/j.fsigen.2013.07.009","article-title":"Population Data of the 46 Insertion\u2013Deletion (INDEL) Loci in Population in Piau\u00ed State, Northeastern Brazil","volume":"9","author":"Lopes","year":"2014","journal-title":"Forensic Sci. Int. Genet."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/16\/4\/372\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T16:59:29Z","timestamp":1760029169000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/16\/4\/372"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,3,24]]},"references-count":18,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2025,4]]}},"alternative-id":["genes16040372"],"URL":"https:\/\/doi.org\/10.3390\/genes16040372","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,3,24]]}}}