Gold nanoprobe functionalized with specific fusion protein selection from phage display and its application in rapid, selective and sensitive colorimetric biosensing of Staphylococcus aureus.

Staphylococcus aureus (S. aureus) is one of the most ubiquitous pathogens in public healthcare worldwide. It holds great insterest in establishing robust analytical method for S. aureus. Herein, we report a S. aureus-specific recognition element, isolated from phage monoclone GQTTLTTS, which was selected from f8/8 landscape phage library against S. aureus in a high-throughput way. By functionalizing cysteamine (CS)-stabilized gold nanoparticles (CS-AuNPs) with S. aureus-specific pVIII fusion protein (fusion-pVIII), a bifunctional nanoprobe (CS-AuNPs@fusion-pVIII) for S. aureus was developed. In this strategy, the CS-AuNPs@fusion-pVIII could be induced to aggregate quickly in the presence of target S. aureus, resulting in a rapid colorimetric response of gold nanoparticles. More importantly, the as-designed probe exhibited excellent selectivity over other bacteria. Thus, the CS-AuNPs@fusion-pVIII could be used as the indicator of target S. aureus. This assay can detect as low as 19CFUmL(-1)S. aureus within 30min. Further, this approach can be applicable to detect S. aureus in real water samples. Due to its sensitivity, specificity and rapidness, this proposed method is promising for on-site testing of S. aureus without using any costly instruments.

[1]  M. Widdowson,et al.  Foodborne Illness Acquired in the United States—Major Pathogens , 2011, Emerging infectious diseases.

[2]  Chunyan Sun,et al.  A novel colorimetric aptasensor using cysteamine-stabilized gold nanoparticles as probe for rapid and specific detection of tetracycline in raw milk , 2015 .

[3]  Chuanbin Mao,et al.  Nanocomposite Films Assembled from Genetically Engineered Filamentous Viruses and Gold Nanoparticles: Nanoarchitecture‐ and Humidity‐Tunable Surface Plasmon Resonance Spectra , 2009 .

[4]  S. Pawar,et al.  Innovative Developments in Bacterial Detection with Magnetic Nanoparticles , 2015, Applied Biochemistry and Biotechnology.

[5]  V. Petrenko,et al.  A Label-Free Electrochemical Impedance Cytosensor Based on Specific Peptide-Fused Phage Selected from Landscape Phage Library , 2016, Scientific Reports.

[6]  Xingyu Jiang,et al.  Gold nanoparticles for the colorimetric and fluorescent detection of ions and small organic molecules. , 2011, Nanoscale.

[7]  B. Åsvold,et al.  Epidemiology and outcome of Staphylococcus aureus bloodstream infection and sepsis in a Norwegian county 1996–2011: an observational study , 2015, BMC Infectious Diseases.

[8]  Thomas Thundat,et al.  Impedimetric detection of pathogenic Gram-positive bacteria using an antimicrobial peptide from class IIa bacteriocins. , 2014, Analytical chemistry.

[9]  V. Petrenko,et al.  Landscape phage fusion protein-mediated targeting of nanomedicines enhances their prostate tumor cell association and cytotoxic efficiency. , 2010, Nanomedicine : nanotechnology, biology, and medicine.

[10]  Dan Zhong,et al.  Employing carbon dots modified with vancomycin for assaying Gram-positive bacteria like Staphylococcus aureus. , 2015, Biosensors & bioelectronics.

[11]  Yaqi Cai,et al.  Sensitive colorimetric visualization of perfluorinated compounds using poly(ethylene glycol) and perfluorinated thiols modified gold nanoparticles. , 2014, Analytical chemistry.

[12]  Olivier Lazcka,et al.  Pathogen detection: a perspective of traditional methods and biosensors. , 2007, Biosensors & bioelectronics.

[13]  R Balfour Sartor,et al.  Microbial influences in inflammatory bowel diseases. , 2008, Gastroenterology.

[14]  W. Qin,et al.  Potentiometric aptasensing of Listeria monocytogenes using protamine as an indicator. , 2014, Analytical chemistry.

[15]  Julian Parkhill,et al.  Evolution of MRSA During Hospital Transmission and Intercontinental Spread , 2010, Science.

[16]  Yu-Chie Chen,et al.  Using biofunctionalized nanoparticles to probe pathogenic bacteria. , 2004, Analytical chemistry.

[17]  Shui-Tong Lee,et al.  A facile assay for direct colorimetric visualization of lipopolysaccharides at low nanomolar level , 2012, Nano Research.

[18]  V. Petrenko,et al.  Liposomes targeted by fusion phage proteins. , 2009, Nanomedicine : nanotechnology, biology, and medicine.

[19]  D. Gillet,et al.  Design and synthesis of single-nanoparticle optical biosensors for imaging and characterization of single receptor molecules on single living cells. , 2007, Analytical chemistry.

[20]  V. Petrenko,et al.  Delivery of siRNA into breast cancer cells via phage fusion protein-targeted liposomes. , 2011, Nanomedicine : nanotechnology, biology, and medicine.

[21]  Langone Jj.  Protein A of Staphylococcus aureus and related immunoglobulin receptors produced by streptococci and pneumonococci. , 1982 .

[22]  G. P. Smith,et al.  A library of organic landscapes on filamentous phage. , 1996, Protein engineering.

[23]  D. Pang,et al.  One-step sensitive detection of Salmonella typhimurium by coupling magnetic capture and fluorescence identification with functional nanospheres. , 2013, Analytical chemistry.

[24]  V. Petrenko,et al.  Targeted delivery of siRNA into breast cancer cells via phage fusion proteins. , 2013, Molecular pharmaceutics.

[25]  Kemin Wang,et al.  A combination of positive dielectrophoresis driven on-line enrichment and aptamer-fluorescent silica nanoparticle label for rapid and sensitive detection of Staphylococcus aureus. , 2015, The Analyst.

[26]  Christopher T. Walsh,et al.  Antibiotics for Emerging Pathogens , 2009, Science.

[27]  V. Petrenko,et al.  Diagnostic probes for Bacillus anthracis spores selected from a landscape phage library. , 2004, Clinical chemistry.

[28]  G. P. Smith,et al.  Filamentous fusion phage: novel expression vectors that display cloned antigens on the virion surface. , 1985, Science.

[29]  Bing Xu,et al.  Using biofunctional magnetic nanoparticles to capture vancomycin-resistant enterococci and other gram-positive bacteria at ultralow concentration. , 2003, Journal of the American Chemical Society.

[30]  A. Somboonkaew,et al.  SPR-DNA array for detection of methicillin-resistant Staphylococcus aureus (MRSA) in combination with loop-mediated isothermal amplification. , 2015, Biosensors & bioelectronics.

[31]  Chuanbin Mao,et al.  Virus-mimetic cytoplasm-cleavable magnetic/silica nanoclusters for enhanced gene delivery to mesenchymal stem cells. , 2013, Angewandte Chemie.

[32]  Z. Su,et al.  L-cysteine functionalized gold nanoparticles for the colorimetric detection of Hg2+ induced by ultraviolet light , 2010, Nanotechnology.

[33]  Salvador Alegret,et al.  Magneto immunoseparation of pathogenic bacteria and electrochemical magneto genosensing of the double-tagged amplicon. , 2009, Analytical chemistry.

[34]  Yu-Chie Chen,et al.  Detection of Staphylococcus aureus by functional gold nanoparticle-based affinity surface-assisted laser desorption/ionization mass spectrometry. , 2015, Analytical chemistry.

[35]  V. Petrenko,et al.  Specific probe selection from landscape phage display library and its application in enzyme-linked immunosorbent assay of free prostate-specific antigen. , 2014, Analytical chemistry.

[36]  Ismail Hakki Boyaci,et al.  SERS-based sandwich immunoassay using antibody coated magnetic nanoparticles for Escherichia coli enumeration. , 2011, The Analyst.

[37]  Bing Xu,et al.  Combining Fluorescent Probes and Biofunctional Magnetic Nanoparticles for Rapid Detection of Bacteria in Human Blood , 2006 .

[38]  Baoxin Li,et al.  A simple and sensitive method for visual detection of heparin using positively-charged gold nanoparticles as colorimetric probes. , 2011, Chemical communications.

[39]  V. Petrenko,et al.  Phages from landscape libraries as substitute antibodies. , 2000, Protein engineering.

[40]  P. Low,et al.  Label-free detection of Staphylococcus aureus captured on immutable ligand arrays. , 2013, ACS applied materials & interfaces.

[41]  Yamei Gao,et al.  Identification and evaluation of a novel peptide binding to the cell surface of Staphylococcus aureus. , 2013, Microbiological research.

[42]  V. Petrenko,et al.  Bio-mimetic Nanostructure Self-assembled from Au@Ag Heterogeneous Nanorods and Phage Fusion Proteins for Targeted Tumor Optical Detection and Photothermal Therapy , 2014, Scientific Reports.

[43]  I-Hsuan Chen,et al.  Landscape phage probes for Salmonella typhimurium. , 2005, Journal of microbiological methods.

[44]  V. Petrenko,et al.  Phagemid vectors for phage display: properties, characteristics and construction. , 2012, Journal of molecular biology.

[45]  M. Los,et al.  Antibody modified gold nanoparticles for fast and selective, colorimetric T7 bacteriophage detection. , 2014, Bioconjugate chemistry.

[46]  Irshad Hussain,et al.  Rational and combinatorial design of peptide capping ligands for gold nanoparticles. , 2004, Journal of the American Chemical Society.

[47]  V. Petrenko,et al.  Peptide microarray with ligands at high density based on symmetrical carrier landscape phage for detection of cellulase. , 2014, Analytical chemistry.

[48]  Helinor Johnston,et al.  Exploitation of Nanotechnology for the Monitoring of Waterborne Pathogens: State-of-the-Art and Future Research Priorities. , 2015, Environmental science & technology.

[49]  V. Petrenko,et al.  Specific ligands for classical swine fever virus screened from landscape phage display library. , 2014, Antiviral research.

[50]  V A Petrenko,et al.  Phage protein‐targeted cancer nanomedicines , 2014, FEBS letters.

[51]  Chuanbin Mao,et al.  Virus-based chemical and biological sensing. , 2009, Angewandte Chemie.

[52]  Xinyong Li,et al.  L-cysteine-modified gold nanostars for SERS-based copper ions detection in aqueous media. , 2014, Langmuir : the ACS journal of surfaces and colloids.