title: Nano-biosensor development for bacterial detection during human kidney infection: use of glycoconjugate-specific antibody-bound gold NanoWire arrays (GNWA).
abstract: Infectious disease, commonly caused by bacterial pathogens, is now the world's leading cause of premature death and third overall cause behind cardiovascular disease and cancer. Urinary Tract Infection (UTI), caused by E. coli bacteria, is a very common bacterial infection, a majority in women (85%) and may result in severe kidney failure if not detected quickly. Among hundreds of strains the bacteria, E. coli 0157:H7, is emerging as the most aggressive one because of its capability to produce a toxin causing hemolytic uremic syndrome (HUS) resulting in death, especially in children. In the present study, a project has been undertaken for developing a rapid method for UTI detection in very low bacteria concentration, applying current knowledge of nano-technology. Experiments have been designed for the development of biosensors using nano-fabricated structures coated with elements such as gold that have affinity for biomolecules. A biosensor is a device in which a biological sensing element is either intimately connected to or integrated within a transducer. The basic principle for the detection procedure of the infection is partly based on the enzyme-linked immunosorbent assay system. Anti-E. coli antibody-bound Gold Nanowire Arrays (GNWA) prepared on anodized porous alumina template is used for the primary step followed by binding of the bacteria containing specimen. An alkaline phosphatase-conjugated second antibody is then added to the system and the resultant binding determined by both electrochemical and optical measurements. Various kinds of GNWA templates were used in order to determine the one with the best affinity for antibody binding. In addition, an efficient method for enhanced antibody binding has been developed with the covalent immobilization of an organic linker Dithiobissuccinimidylundecanoate (DSU) on the GNWA surface. Studies have also been conducted to optimize the antibody-binding conditions to the linker-attached GNWA surfaces for their ability to detect bacteria in clinical concentrations.
authors.affiliation: Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, IN, 46556, USA.
authors.forename: Manju
authors.initials: M
authors.lastname: Basu
authors.forename: Sara
authors.initials: S
authors.lastname: Seggerson
authors.forename: Joshua
authors.initials: J
authors.lastname: Henshaw
authors.forename: Juan
authors.initials: J
authors.lastname: Jiang
authors.forename: Rocio
authors.initials: R
authors.lastname: del A Cordona
authors.forename: Clare
authors.initials: C
authors.lastname: Lefave
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Boyle
authors.forename: Albert
authors.initials: A
authors.lastname: Miller
authors.forename: Michael
authors.initials: M
authors.lastname: Pugia
authors.forename: Subhash
authors.initials: S
authors.lastname: Basu
mesh_terms: D000907:Antibodies, Bacterial; D015374:Biosensing Techniques / Q000295:instrumentation*; D004926:Escherichia coli / Q000302:isolation & purification*; D004927:Escherichia coli Infections / Q000175:diagnosis*; D006046:Gold; D006801:Humans; D036103:Nanotechnology / Q000295:instrumentation* / Q000379:methods; D013057:Spectrum Analysis; D014552:Urinary Tract Infections / Q000382:microbiology
chemical_list: D000907:Antibodies, Bacterial; D006046:Gold