Abstract
The timely and accurate diagnosis of infectious diseases is one of the greatest challenges currently facing modern medicine. The development of innovative techniques for the rapid and accurate identification of bacterial pathogens in point-of-care facilities using low-cost, portable instruments is essential. We have developed a novel all-electronic biosensor that is able to identify bacteria in less than ten minutes. This technology exploits bacteriocins, protein toxins naturally produced by bacteria, as the selective biological detection element. The bacteriocins are integrated with an array of potassium-selective sensors in Complementary Metal Oxide Semiconductor technology to provide an inexpensive bacterial biosensor. An electronic platform connects the CMOS sensor to a computer for processing and real-time visualization. We have used this technology to successfully identify both Gram-positive and Gram-negative bacteria commonly found in human infections.
Highlights
The World Health Organization reports that infectious diseases cause 26% of all deaths globally, and account for 45% of the global disease burden [1]
Each strain was grown in 50 mL of LB-Lennox Broth (Sigma Aldrich) at 37°C with shaking to an optical density at 600 nm (OD600) of 0.8, at which point mitomycin C was added to a final concentration of 1μg/mL to induce colicin production and the cells were incubated for 3.5 hours
We examined the ability of pore-forming colicins A9, E1, K, and B [8, 11,12,13] to elicit efflux of potassium from E. coli using a commercially available potassium-selective electrode
Summary
The World Health Organization reports that infectious diseases cause 26% of all deaths globally, and account for 45% of the global disease burden [1]. Electrochemical sensors have been used extensively in biosensors employing DNA as the recognition element, but these tests require significant preparatory steps, including lysing the pathogen and extracting and amplifying the bacterial DNA. As a result, these assays typically take several hours to perform and usually do not distinguish between live and dead bacteria. When a sample containing bacteria is mixed with a bacteriocin to which it is sensitive, a pore is formed in the bacterial cell envelope and potassium efflux occurs from the interior of the cell into the bulk medium (Fig 1a) This phenomenon can be monitored by an ion-selective probe and provides a rapid method for bacterial typing. We leverage extensive integration of the biosensor platform in inexpensive, nanometer scale ultra-small Complementary Metal Oxide Semiconductor (CMOS) technology along with low-noise reading circuitry to provide a low-cost, portable biosensor able to accurately detect and identify bacteria in less than ten minutes
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