Abstract
A novel dual-peptide recognition strategy was designed for sandwich fluorimetric detection of Staphylococcus aureus (S. aureus) utilizing the affinity of bacterial cells for certain peptides. A phage-display peptide derived from a random peptide library was used to functionalize magnetic particles and to specifically capture S. aureus. Magainin I is a broad-spectrum antimicrobial peptide that binds to cell membranes of most bacteria. It was used as a second recognition peptide to form a sandwich complex with S. aureus. By using fluorescein isothiocyanate as the fluorescent label and working at excitation/emission wavelengths of 488/525 nm, S. aureus can be directly detected in the 10 to 10,000 cfu·mL−1 concentration range, with a detection limit as low as 9 cfu·mL−1. The whole detection process can be completed within 90 min. This strategy was successfully applied to the detection of S. aureus in spiked lake water, human urine and apple juice. Respective recovery values ranged from 81% to 110%. The assay is highly sensitive and specific, and fast. The use of synthetic peptides shows many advantages such as lower cost, higher stability and ease of chemical modification compared to other molecular recognition agents such as antibodies, bacteriophages, or aptamers. In our perception, this detection scheme may be extended to many other pathogens if phage-displayed peptides specific for other bacteria are available.
References
Rozemeijer W, Fink P, Rojas E, Jones CH, Pavliakova D, Giardina P, Murphy E, Liberator P, Jiang Q, Girgenti D, Peters RP, Savelkoul PH, Jansen KU, Anderson AS, Kluytmans J (2015) Evaluation of approaches to monitor Staphylococcus aureus virulence factor expression during human disease. PLoS One 10:116945
Lowy FD (1998) Staphylococcus aureus Infections. N Engl J Med 339:2025–2027
Paule SM, Mehta M, Hacek DM, Gonzalzles TM, Robicsek A, Peterson LR (2009) Chromogenic media vs real-time PCR for nasal surveillance of methicillin-resistant Staphylococcus aureus impact on detection of MRSA-positive persons. Am J Clin Pathol 131:532–539
Lazcka O, Campo FJD, Muñoz FX (2007) Pathogen detection: a perspective of traditional methods and biosensors. Biosens Bioelectron 22:1205–1217
Kobayashi N, Wu H, Kojima K, Taniguchi K, Urasawa S, Uehara N, Omizu Y, Kishi Y, Yagihashi A, Kurokawa I (1994) Detection of mecA, femA, and femB genes in clinical strains of staphylococci using polymerase chain-reaction. Epidemiol Infect 113:259–266
Wang H, Chen HW, Hupert ML, Chen PC, Pittman PT, Goettert J, Murphy MC, Williams D, Barany F (2012) Fully integrated thermoplastic genosensor for the highly sensitive detection and identification of mulit-drug-resistant tuberculosis. Angew Chem Int Ed 51:4349–4353
Li YW, Yan XH, Feng XJ, Wang J, Du W, Wang YC, Chen P, Xiong L, Liu BF (2014) Agarose-based microfluidic device for point-of-care concentration and detection of pathogen. Anal Chem 86:10653–10659
Bhardwaj N, Bhardwaj SK, Mehta J, Mohanta GC, Deep A (2016) Bacteriophage immobilized graphene electrodes for impedimetric sensing of bacteria (Staphylococcus arlettae). Anal Biochem 505:18–25
Chang YC, Yang CY, Sun RL, Cheng YF, Kao WC, Yang PC (2013) Rapid single cell detection of Staphylococcus aureus by aptamer-conjugated gold nanoparticles. Sci Rep 3:31863–31870
Carrillo-Carrion C, Simonet BM, Valcarcel M (2011) Colistin-functionalised CdSe/ZnS quantum dots as fluorescent probe for the rapid detection of Escherichia coli. Biosens Bioelectron 26:4368–4374
Yu JP, Zhang Y, Zhang Y, Li H, Yang H, Wei HP (2016) Sensitive and rapid detection of Staphylococcus aureus in milk via cell binding domain of lysin. Biosens Bioelectron 77:366–371
Dechtrirat D, Gajovic-Eichlmann N, Wojick F, Hartmann L, Bier FF, Scheller FW (2014) Electrochemical displacement sensor based on ferrocene boronic acid tracer and immobilized glycan for saccharide binding proteins and E. coli. Biosens Bioelectron 58:1–8
Wu SJ, Duan N, Gu HJ, Hao LL, Ye H, Gong WH, Wang ZP (2016) A review of the methods for detection of Staphylococcus aureus enterotoxins. Toxins 8:176–196
Mannoor MS, Zhang S, Link AJ, McAlpine MC (2010) Electrical detection of pathogenic bacteria via immobilized antimicrobial peptides. Proc Natl Acad Sci U S A 107:19207–19212
Kulagina NV, Lassman ME, Ligler FS, Taitt CR (2005) Antimicrobial peptide for detection of bacteria in biosensor assays. Anal Chem 77:6504–6508
Nielsen KM, Kyneb MH, Alstrup AKO, Jensen JJ, Bender D, Schønheyder HC, Afzelius P, Nielsen OL, Jensen SB (2016) 68Ga-labeled phage-display selected peptides as tracers for positron emission tomography imaging of Staphylococcus aureus biofilm-associated infections: selection, radiolabelling and preliminary biological evaluation. Nucl Med Biol 43:593–605
Liu P, Han L, Wang F, Petrenko VA, Liu A (2016) Gold nanoprobe functionalized with specific fusion protein selection from phage display and its application in rapid, selective and sensitive colorimetric biosensing of Staphylococcus aureus. Biosens Bioelectron 15:195–203
Rao SS, Mohan KVK, Gao YM, Atreya CD (2013) Identification and evaluation of a novel peptide binding to the cell surface of Staphylococcus aureus. Mirobiol Res 168:106–112
Costa F, Carvalho IF, Montelaro RC, Gomes P, Martins MCL (2011) Covalent immobilization of antimicrobial peptides (AMPs) onto biomaterial surfaces. Acta Biomater 7:1431–1440
Xue XH, Pan J, Xie HM, Wang JH, Zhang S (2009) Fluorescence detection of total count of Escherichia coli and Staphylococcus aureus on water-soluble CdSe quantum dots coupled with bacteria. Talanta 77:1808–1813
Wang XL, Huang YK, Wu SJ, Duan N, Xu BC, Wang ZP (2016) Simultaneous detection of Staphylococcus aureus and Salmonella typhimurium using multicolor time-resolved fluorescence nanopartilces as lables. Int J Food Microbiol 237:172–179
Zhang H, Ma XY, Liu Y, Duan Nuo WSJ, Wang ZP, Xu BC (2015) Gold nanoparticles enhanced SERS aptasensor for the simultaneous detection of Salmonella typhimurium and Staphylococcus aureus. Biosens Bioelectron 74:872–877
Shojaei TR, Swlleh MAM, Tabatabaei M, Ekrami A, Motallebi R, Rahmani-Cherati T, Hajalilou A, Raheleh J (2014) Development of sandwich-form biosensor to detect Mycobacterium tuberculosis complex in clinical sputum specimens. Braz J Infect Dis 8:600–608
Morton J, Karoonuthaisiri N, Stewart LD, Oplatowska M, Elliott CT, Grant IR (2013) Production and evaluation of the utility of novel phage display-derived peptide ligands to Salmonella spp for magnetic separation. J Appl Microbiol 115:271–281
Rao SS, Mohan KVK, Atreya CD (2013) A peptide derived from phage display library exhibits antibacterial activity against E. coli and Pseudomonas aeruginosa. PloS ONE 2: 56081
Hao LL, Gu HJ, Duan N, Wu SJ, Ma XY, Xia Y, Tao Z, Wang ZP (2017) An enhanced chemiluminescence resonance energy transfer aptasensor based on rolling circle amplification and WS2 nanosheet for Staphylococcus aureus detection. Anal Chim Acta 959:83–90
Xiong J, Wang WW, Zhou YL, Kong WJ, Wang ZX, Fu ZF (2016) Ultra-sensitive chemiluminescent detection of Staphylococcus aureus based on competitive binding of Staphylococcus protein A-modified magnetic beads to immunoglobulin G. Microchim Acta 183:1507–1512
Zhang B, Li HH, Pan WX, Chen QS, Ouyang Q, Zhao JW (2016) Dual-color upconversion nanoparticles (UCNPs)-based fluorescent immunoassay probes for sensitive sensing foodborne pathogens. Food Anal Methods 6:2036–2045
Wang BB, Wang Q, Ma MH, Cai ZX (2015) The inhibition of fluorescence resonance energy transfer between multicolor quantum dots for rapid and sensitive detection of Staphylococcus aureus. Spectrochim Acta 135:428–434
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This work was financially supported by the Natural Science Foundation of China (21475107) and the Fundamental Research Funds for the Central Universities (XDJK2017A008).
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Xiong, J., Wang, W. & Fu, Z. Fluorimetric sandwich affinity assay for Staphylococcus aureus based on dual-peptide recognition on magnetic nanoparticles. Microchim Acta 184, 4197–4202 (2017). https://doi.org/10.1007/s00604-017-2396-8
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DOI: https://doi.org/10.1007/s00604-017-2396-8