Abstract
Pseudomonas aeruginosa has been considered as a representative pathogenic bacteria in drinking water. In order to detect P. aeruginosa, aptamers were utilized in this study. In particular, fluorescein isothiocyannate (FITC) and quantum dot (QD) were used for aptamer labeling. FITC-labeled aptamers showed higher binding capacity with optimal incubation time of 30 min compared to QD-labeled aptamers. However, incubation speed did not have any effect on the binding capacity of FITC-labeled aptamers to bacteria. Aptamer-binding capacity was measured according to varying cell concentrations of 0, 10, 100, and 1000 cells/mL. As a result, the limit of detection, limit of quantification, and limit of linearity of P. aeruginosa were 5.07, 5.64, and 100 cells/mL, respectively. The low detection limit shows the fluorophore-labeled aptamer potential to detect P. aeruginosa labeling in the field.
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References
Akerlund T, Nordström K, and Bernander R (1995) Analysis of cell size and DNA content in exponentially growing and stationary-phase batch cultures of Escherichia coli. J Bacteriol 177, 6791–6797.
Cai J, Yao C, Xia J, Wang J, Chen M, Huang J et al. (2011) Rapid parallelized and quantitative analysis of five pathogenic bacteria by ITS hybridization using QCM biosensor. Sensor Actuat B-Chem 155, 500–504.
Cao X, Li S, Chen L, Ding H, Xu H, Huang Y et al. (2009) Combining use of a panel of ssDNA aptamers in the detection of Staphylococcus aureus. Nucleic Acids Res 37, 4621–4628.
Carnazza S, Foti C, Gioffrè G, Felici F, and Guglielmino S (2008) Specific and selective probes for Pseudomonas aeruginosa from phage-displayed random peptide libraries. Biosens Bioelectron 23, 1137–1144.
Carpani I, Conti P, Lanteri S, Legnani PP, Leoni E, and Tonelli D (2008) Direct quantification of test bacteria in synthetic water-polluted samples by square wave voltammetry and chemometric methods. Biosens Bioelectron 23, 959–964.
Doorneweerd DD, Henne WA, Reifenberger RG, and Low PS (2010) Selective capture and identification of pathogenic bacteria using an immobilized siderophore. Langmuir 26, 15424–15429.
Dwarakanath S, Bruno JG, Shastry A, Phillips T, John A, Kumar A et al. (2004) Quantum dot-antibody and aptamer conjugates shift fluorescence upon binding bacteria. Biochem Bioph Res Co 325, 739–743.
Esiobu N, Mohammed R, Echeverry A, Green M, Bonilla T, Hartz A et al. (2004) The application of peptide nucleic acid probes for rapid detection and enumeration of eubacteria, Staphylococcus aureus and Pseudomonas aeruginosa in recreational beaches of S. Florida. J Microbiol Meth 57, 157–162.
Gilmartin N and O’Kennedy R (2012) Nanobiotechnologies for the detection and reduction of pathogens. Enzyme Microb Tech 50, 87–95.
He F and Liu S (2004) Detection of P. aeruginosa using nano-structured electrode-separated piezoelectric DNA biosensor. Talanta 62, 271–277.
Joshi R, Janagama H, Dwivedi HP, Senthil Kumar TMA, Jaykus LA, Schefers J et al. (2009). Selection, characterization, and application of DNA aptamers for the capture and detection of Salmonella enterica serovars. Mol Cell Probe 23, 20–28.
Kim N, Park I, and Kim D (2004) Characteristics of a label-free piezoelectric immunosensor detecting Pseudomonas aeruginosa. Sensor Actuat BChem 100, 432–438.
Ligler FS (2009) Perspective on optical biosensors and integrated sensor systems. Anal Chem 81, 519–526.
Liu C, Zeng GM, Tang L, Zhang Y, Li YP, Liu YY et al. (2011) Electrochemical detection of Pseudomonas aeruginosa 16S rRNA using a biosensor based on immobilized stem-loop structured probe. Enzyme Microb Tech 49, 266–271.
Mena KD and Gerba CP (2009) Risk assessment of Pseudomonas aeruginosa in water. Rev Environ Contam Toxicol 201, 71–115.
Navarro Liorens JM, Tormo A, and Martinez-Garcia E (2010) Stationary phase in gram-negative bacteria. FEMS Microbiol Rev 34, 476–495.
Pang P, Huang S, Cai Q, Yao S, Zeng K, and Grimes CA (2007) Detection of Pseudomonas aeruginosa using a wireless magnetoelastic sensing device. Biosens Bioelectron 23, 295–299.
Paoli GC (2006) Molecular approaches to immunological biosensors: phage displayed antibodies for the detection of foodborne pathogenic bacteria. In Advances in Microbial Food Safety (Vol. 931), pp. 41–54, ACS publications, Washington, DC, USA.
Park JS, Lee CM, and Lee KY (2007) A surface plasmon resonance biosensor for detecting Pseudomonas aeruginosa cells with self-assembled chitosan-alginate multilayers. Talanta 72, 859–862.
Reinemann C, Stoltenburg R, and Strehlitz B (2009) Investigations on the specificity of DNA aptamers binding to ethanolamine. Anal Chem 81, 3973–3978.
Rodriguez-Mozaz S, Lopez de Alda MJ, and Barceló D (2006) Biosensors as useful tools for environmental analysis and monitoring. Anal Bioanal Chem 386, 1025–1041.
Rusin PA, Rose JB, Haas CN, and Gerba CP (1997) Risk assessment of opportunistic bacterial pathogens in drinking water. Rev Environ Contam Toxicol 152, 57–83.
Sassolas A, Leca-Bouvier BD, and Blum LJ (2008) DNA biosensors and microarrays. Chem Rev 108, 109–139.
Song S, Wang L, Li J, Fan C, and Zhao J (2008) Aptamer-based biosensors. TrAC-Trend Anal Chem 27, 108–117.
Tennico YH, Hutanu D, Koesdjojo MT, Bartel CM, and Remcho VT (2010) On-chip aptamer-based sandwich assay for thrombin detection employing magnetic beads and quantum dots. Anal Chem 82, 5591–5597.
Torres-Chavolla E, and Alocilja EC (2009) Aptasensors for detection of microbial and viral pathogens. Biosens Bioelectron 24, 3175–3182.
Wang KY, Zeng YL, Yang XY, Li WB, and Lan XP (2011) Utility of aptamer-fluorescence in situ hybridization for rapid detection of Pseudomonas aeruginosa. Eur J Clin Microbiol Infect Dis 30, 273–278.
Wang Y, Hammes F, Düggelin M, and Egli T (2008) Influence of size, shape, and flexibility on bacterial passage through micropore membrane filters. Environ Sci Technol 42, 6749–6754.
Zhao Y, Ye M, Chao Q, Jia N, Ge Y, and Shen H (2009) Simultaneous detection of multifood-borne pathogenic bacteria based on functionalized quantum dots coupled with immunomagnetic separation in food samples. J Agr Food Chem 57, 517–524.
Zhou G, Wen S, Liu Y, Li R, Zhong X, Feng L et al. (2011) Development of a DNA microarray for detection and identification of Legionella pneumophila and ten other pathogens in drinking water. Int J Food Microbiol 145, 293–300.
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Kim, L.H., Yu, HW., Kim, YH. et al. Potential of fluorophore labeled aptamers for Pseudomonas aeruginosa detection in drinking water. J Korean Soc Appl Biol Chem 56, 165–171 (2013). https://doi.org/10.1007/s13765-013-3019-7
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DOI: https://doi.org/10.1007/s13765-013-3019-7