Optical and electrochemical detection of a verotoxigenic E. coli gene using DNAzyme-labeled stem-loops
Autor: | Gloria Longinotti, Gabriel Ybarra, Javier Marcelo Montserrat |
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Jazyk: | angličtina |
Rok vydání: | 2017 |
Předmět: |
Genosensors
Genómica ADN Deoxyribozyme DNAzyme Peroxidase 02 engineering and technology 010402 general chemistry Electrochemistry biosensor 01 natural sciences Buffer (optical fiber) Electrochemical cell lcsh:Chemistry purl.org/becyt/ford/1 [https] Verotoxigenic E. coli Colloid and Surface Chemistry Biosensores DNAzyme peroxidase Materials Chemistry purl.org/becyt/ford/1.4 [https] Chemical Engineering (miscellaneous) Detection limit Escherichia Coli Chromatography Métodos ópticos Chemistry Otras Ciencias Químicas Biosensors Ciencias Químicas Métodos electroquímicos 021001 nanoscience & nanotechnology Detección 0104 chemical sciences lcsh:QD1-999 Electrode 0210 nano-technology Selectivity Biosensor vertoxigenic E. coli CIENCIAS NATURALES Y EXACTAS genosensor |
Zdroj: | Journal of Electrochemical Science and Engineering Volume 7 Issue 4 Journal of Electrochemical Science and Engineering, 7(4) Repositorio Institucional del Instituto Nacional de Tecnología Industrial (INTI) Instituto Nacional de Tecnología Industrial instacron:INTI CONICET Digital (CONICET) Consejo Nacional de Investigaciones Científicas y Técnicas instacron:CONICET Journal of Electrochemical Science and Engineering, Vol 7, Iss 4 (2017) |
ISSN: | 1847-9286 |
Popis: | The activity of a peroxidase-mimicking DNAzyme was optimized to be used as a catalytic label in a stem-loop genosensor construction for quantifying the gene sequence Shiga-like toxin I of verotoxigenic E. coli. Experimental conditions such as pH, buffer composition, potassium ion concentration, and hemin-to-oligonucleotides ratio, were analyzed to maximize optical and electrochemical responses using microvolumes. Different stem-loop constructions were evaluated to obtain the optimum response against the target concentration. Linear ranges of 0.05-0.5 µM and limits of detection of 174 nM and 144 nM were estimated for the optical and electrochemical measurements, respectively. Selectivity was proved by assaying other verotoxigenic, enterotoxigenic and enteroinvasive sequences. The results show that, if a combination of small-volume electrochemical cells and low-cost untreated screen-printed electrodes with a relatively high geometric area is used, electrochemical measurements present similar sensitivity and limits of detection to the more usual optical ones, allowing the development of low-cost electrochemical biosensors based on the use of soluble DNAzymes as labels. Fil: Longinotti, Gloria. Instituto Nacional de Tecnología Industrial; Argentina Fil: Ybarra, Gabriel Omar. Instituto Nacional de Tecnología Industrial; Argentina Fil: Montserrat, Javier Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de General Sarmiento; Argentina |
Databáze: | OpenAIRE |
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