Osteoporosis is a multifactorial disease influenced by genetic and environmental factors, which contributes to an increased risk of bone fracture, but early diagnosis of this disease cannot be achieved using current techniques. We describe a generic platform for the targeted electrochemical genotyping of SNPs identified by genome-wide association studies to be associated with a genetic predisposition to osteoporosis. The platform exploits isothermal solid-phase primer elongation with ferrocene-labeled nucleoside triphosphates. Thiolated reverse primers designed for each SNP were immobilized on individual gold electrodes of an array. These primers are designed to hybridize to the SNP site at their 3'OH terminal, and primer elongation occurs only where there is 100% complementarity, facilitating the identification and heterozygosity of each SNP under interrogation. The platform was applied to real blood samples, which were thermally lysed and directly used without the need for DNA extraction or purification. The results were validated using Taqman SNP genotyping assays and Sanger sequencing. The assay is complete in just 15 min with a total cost of 0.3€ per electrode. The platform is completely generic and has immense potential for deployment at the point of need in an automated device for targeted SNP genotyping with the only required end-user intervention being sample addition.

Department of Internal Medicine Erasmus MC 40 3015 Rotterdam The Netherlands

Department of Organic Chemistry Faculty of Science Charles University Hlavova 8 CZ 12843 Prague 2 Czech Republic

Division of Endocrinology and Diabetology Department of Internal Medicine Medical University of Graz 8036 Graz Austria

Institució Catalana de Recerca i Estudis Avancats 08010 Barcelona Spain

Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences Flemingovo namesti 2 CZ 16610 Prague 6 Czech Republic

INTERFIBIO Research Group Departament d'Enginyeria Química Universitat Rovira i Virgili 43007 Tarragona Spain

Labman Automation Ltd Seamer Hill Stokesley North Yorkshire TS9 5NQ U K

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