Electrochemical primer extension based on polyoxometalate electroactive labels for multiplexed detection of single nucleotide polymorphisms
| Autor: | Serge Thorimbert, Bernold Hasenknopf, Ahmed M. Debela, Ciara K. O'Sullivan, Ioannis Katakis, Laura A. Uribe, Nassif Chahin, Mayreli Ortiz |
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| Přispěvatelé: | Departament d′Enginyeria Quimica, Universitat Rovira i Virgili, Departament d′Enginyeria Quıímica, Institut Parisien de Chimie Moléculaire (IPCM), Chimie Moléculaire de Paris Centre (FR 2769), Institut de Chimie du CNRS (INC)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institució Catalana de Recerca i Estudis Avançats (ICREA) |
| Rok vydání: | 2018 |
| Předmět: |
Biomedical Engineering
Biophysics [CHIM.THER]Chemical Sciences/Medicinal Chemistry Biosensing Techniques 010402 general chemistry Electrochemistry Polymorphism Single Nucleotide 01 natural sciences Primer extension Nucleobase chemistry.chemical_compound Humans Peptide bond [CHIM.COOR]Chemical Sciences/Coordination chemistry ComputingMilieux_MISCELLANEOUS DNA Primers [CHIM.ORGA]Chemical Sciences/Organic chemistry 010405 organic chemistry Hybridization probe Reproducibility of Results General Medicine Tungsten Compounds 0104 chemical sciences Crystallography chemistry Electrode Polyoxometalate [CHIM.CHEM]Chemical Sciences/Cheminformatics DNA Biotechnology |
| Zdroj: | Biosensors and Bioelectronics Biosensors and Bioelectronics, Elsevier, 2018, 117, pp.201-206. ⟨10.1016/j.bios.2018.06.014⟩ |
| ISSN: | 0956-5663 |
| Popis: | Polyoxymetalates (POMs) ([SiW 11 O 39 {Sn(CH 2 ) 2 CO)}] 4- and [P 2 W 17 O 61 {Sn(CH 2 ) 2 CO)}] 6- ) were used to modify dideoxynucleotides (ddNTPs) through amide bond formation, and applied to the multiplexed detection of single nucleotide polymorphisms (SNPs) in an electrochemical primer extension reaction. Each gold electrode of an array was functionalised with a short single stranded thiolated DNA probe, specifically designed to extend with the POM-ddNTP at the SNP site to be interrogated. The system was applied to the simultaneous detection of 4 SNPs within a single stranded 103-mer model target generated using asymmetric PCR, highlighting the potential of POM-ddNTPs for targeted, multiplexed SNP detection. The four DNA bases were successfully labelled with both ([SiW 11 O 39 {Sn(CH 2 ) 2 CO)}] 4- and [P 2 W 17 O 61 {Sn(CH 2 ) 2 CO)}] 6- ), and [SiW 11 O 39 {Sn(CH 2 ) 2 CO)}] 4- demonstrated to be the more suitable due to its single oxidation peak, which provides an unequivocal signal. The POM-ddNTP enzymatically incorporated to the DNA anchored to the surface was visualised by AFM using gold coated mica. The developed assay has been demonstrated to be highly reproducible, simple to carry out and with very low non-specific background signals. Future work will focus on applying the developed platform to the detection of SNPs associated with rifampicin resistance in real samples from patients suffering from tuberculosis. |
| Databáze: | OpenAIRE |
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