A fluorescence quenching based gene assay for Escherichia coli O157:H7 using graphene quantum dots and gold nanoparticles
| Autor: | Jaafar Abdullah, Suria Mohd Saad, Yap Wing Fen, Suraya Abd Rashid, Lau Han Yih, Faridah Salam |
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| Rok vydání: | 2019 |
| Předmět: |
DNA
Bacterial Fluorophore Immobilized Nucleic Acids Metal Nanoparticles Food Contamination 02 engineering and technology Escherichia coli O157 010402 general chemistry medicine.disease_cause 01 natural sciences Poultry Analytical Chemistry chemistry.chemical_compound Cheese Quantum Dots Spectroscopy Fourier Transform Infrared medicine Animals Escherichia coli Detection limit Quenching (fluorescence) Oligonucleotide Escherichia coli Proteins Nucleic Acid Hybridization 021001 nanoscience & nanotechnology Fluorescence 0104 chemical sciences chemistry Colloidal gold Biophysics Graphite Gold 0210 nano-technology DNA Flagellin |
| Zdroj: | Microchimica Acta. 186 |
| ISSN: | 1436-5073 0026-3672 |
| DOI: | 10.1007/s00604-019-3913-8 |
| Popis: | A fluorometric assay is described for highly sensitive quantification of Escherichia coli O157:H7. Reporter oligos were immobilized on graphene quantum dots (GQDs), and quencher oligos were immobilized on gold nanoparticles (AuNPs). Target DNA was co-hybridized with reporter oligos on the GQDs and quencher oligos on AuNPs. This triggers quenching of fluorescence (with excitation/emission peaks at 400 nm/530 nm). On introducing target into the system, fluorescence is quenched by up to 95% by 100 nM concentrations of target oligos having 20 bp. The response to the fliC gene of E. coli O157:H7 increases with the logarithm of the concentration in the range from 0.1 nM to 150 nM. The limit of detection is 1.1 ± 0.6 nM for n = 3. The selectivity and specificity of the assay was confirmed by evaluating the various oligos sequences and PCR product (fliC gene) amplified from genomic DNA of the food samples spiked with E. coli O157:H7. Graphical abstractSchematic representation of fluorometric assay for highly sensitive quantification of Escherichia coli O157:H7 based on fluorescence quenching gene assay for fliC gene of E. coli O157:H7. |
| Databáze: | OpenAIRE |
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