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Enzyme-free hybridization chain reaction-based signal amplification strategy for the sensitive detection of Staphylococcus aureus.
- Source :
-
Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy [Spectrochim Acta A Mol Biomol Spectrosc] 2019 May 15; Vol. 215, pp. 41-47. Date of Electronic Publication: 2019 Feb 19. - Publication Year :
- 2019
-
Abstract
- In this study, based on hybridization chain reaction (HCR) amplification and graphene oxide (GO), we developed a facile enzyme-free signal amplification strategy for sensitive detection of Staphylococcus aureus (S. aureus). Two hairpin probes (HP <subscript>1</subscript> and HP <subscript>2</subscript> ) labeled by fluorophore 6-carboxyfluorescein (FAM) are designed. The HP <subscript>1</subscript> and HP <subscript>2</subscript> can not only trigger to the HCR but also form a long nicked double strand DNA (dsDNA) with the target (16S rRNA). In the absence of target (16 s RNA), the free FAM-labeled HP1 and HP2 are adsorbed by the GO via π-π stacking, the fluorescence signal is quenched. In the presence of target (16 s RNA), the HCR is triggered and dsDNA complexes are generated. As a result, the fluorescence signal can be strongly amplified by the synergistic effect of FAM and the dsDNA dye SYBR Green I. Based on this mechanism, a fluorescence method is designed for the detection of 16S rRNA of S. aureus. Under the optimal conditions, it has low detection limit (50 pM) and a linear response in a concentration range of 50 pM to 100 nM for 16S rRNA. Furthermore, this method has also been successfully applied to the detection of S. aureus in milk sample with the detection limit of 4 × 10 <superscript>2</superscript> CFU·mL <superscript>-1</superscript> .<br /> (Copyright © 2019 Elsevier B.V. All rights reserved.)
- Subjects :
- Animals
Benzothiazoles
Biosensing Techniques
DNA, Bacterial analysis
Diamines
Fluoresceins
Fluorescent Dyes chemistry
Graphite chemistry
Limit of Detection
Milk microbiology
Organic Chemicals
Quinolines
Food Microbiology methods
Molecular Typing methods
Nucleic Acid Hybridization methods
Staphylococcus aureus isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1873-3557
- Volume :
- 215
- Database :
- MEDLINE
- Journal :
- Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy
- Publication Type :
- Academic Journal
- Accession number :
- 30818216
- Full Text :
- https://doi.org/10.1016/j.saa.2019.02.035