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.

Use of Electrochemical DNA Biosensors for Rapid Molecular Identification of Uropathogens in Clinical Urine Specimens

“…Amperometric biosensors for detection of in vitro cultivated bacteria using antibody capture or nucleic acid hybridization approaches have been described, but not for detection of bacteria in clinical specimens (1,4,9,10,15,21,26,30,35,40,43). A convergence of technological innovations from several disciplines, including microfabrication, materials science, electrochemistry, and molecular microbiology, contributed to the design of the electrochemical sensor array and the detection strategy utilized in the current study (16,17,42).…”

Current status of nanotechnology in urology

“…Based on both optical and electrochemical nanoparticles and specific E. coli antibodies, a quick and sensitive procedure has been developed for bacterial detection in case of kidney infection. 11 The study showed that the biosensor can detect each of fifty E. coli cells with the sensor area of 0.178 cm 2 . Nanosensor based microbial detection will not only improve diagnostic accuracy but also cure rates.…”

Materiomics for Oral Disease Diagnostics and Personal Health Monitoring: Designer Biomaterials for the Next Generation Biomarkers

“…For example, some metal nanowires can be adopted in the detection of glucose (Cusma et al, 2007;Lu et al, 2007;Qu et al, 2007), cholesterol (Aravamudhan et al, 2007a;2007b), or DNA (Gao et al, 2007;Yi et al, 2010). Other types of nanowires have been exploited to detect inosine (Liu et al, 2006), bacterial (Basu et al, 2004;Garcia-Aljaro et al, 2010;Mishra et al, 2008), or cancer biomarkers (Choi et al, 2010;Ramgir et al, 2007;Zheng et al, 2005).…”