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Generic Platform for the Multiplexed Targeted Electrochemical Detection of Osteoporosis-Associated Single Nucleotide Polymorphisms Using Recombinase Polymerase Solid-Phase Primer Elongation and Ferrocene-Modified Nucleoside Triphosphates
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SYSNO ASEP 0574261 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Generic Platform for the Multiplexed Targeted Electrochemical Detection of Osteoporosis-Associated Single Nucleotide Polymorphisms Using Recombinase Polymerase Solid-Phase Primer Elongation and Ferrocene-Modified Nucleoside Triphosphates Author(s) Ortiz, M. (ES)
Jauset-Rubio, M. (ES)
Trummer, O. (AT)
Foessl, I. (AT)
Kodr, David (UOCHB-X) ORCID
Acero, J. L. (ES)
Botero, M. L. (ES)
Biggs, P. (GB)
Lenartowicz, D. (GB)
Trajanoska, K. (NL)
Rivadeneira, F. (NL)
Hocek, Michal (UOCHB-X) RID, ORCID
Obermayer-Pietsch, B. (AT)
O'Sullivan, C. K. (ES)Source Title ACS Central Science. - : American Chemical Society - ISSN 2374-7943
Roč. 9, č. 8 (2023), s. 1591-1602Number of pages 12 s. Language eng - English Country US - United States Keywords bone mineral density ; chain reaction ; DNA OECD category Organic chemistry R&D Projects GX20-00885X GA ČR - Czech Science Foundation (CSF) Method of publishing Open access Institutional support UOCHB-X - RVO:61388963 UT WOS 001032190400001 EID SCOPUS 85166762969 DOI 10.1021/acscentsci.3c00243 Annotation 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. Workplace Institute of Organic Chemistry and Biochemistry Contact asep@uochb.cas.cz ; Kateřina Šperková, Tel.: 232 002 584 ; Jana Procházková, Tel.: 220 183 418 Year of Publishing 2024 Electronic address https://doi.org/10.1021/acscentsci.3c00243
Number of the records: 1