title: Rapid detection of Escherichia coli O157:H7 and Salmonella Typhimurium in foods using an electrochemical immunosensor based on screen-printed interdigitated microelectrode and immunomagnetic separation.
abstract: Foodborne pathogens have continuously been a serious food safety issue and there is a growing demand for a rapid and sensitive method to screen the pathogens for on-line or in-field applications. Therefore, an impedimetric immunosensor based on the use of magnetic beads (MBs) for separation and a screen-printed interdigitated microelectrode (SP-IDME) for measurement was studied for the rapid detection of Escherichia coli O157:H7 and Salmonella Typhimurium in foods. Streptavidin coated MBs were functionalized with corresponding biotinylated antibodies (Ab) to capture the target bacteria. The glucose oxidase (GOx)-Ab conjugates were employed to label the MBs-Ab-cell complexes. The yielded MBs-Ab-cell-Ab-GOx biomass was mixed with the glucose solution to trigger an enzymatic reaction which produced gluconic acid. This increased the ion strength of the solution, thus decreasing the impedance of the solution measured on the SP-IDME. Our results showed that the immunosensor was capable of specifically detecting E. coli O157:H7 and S. Typhimurium within the range of 10(2)-10(6) cfu ml(-1) in the pure culture samples. E. coli O157:H7 in ground beef and S. Typhimurium in chicken rinse water were also examined. The limits of detection (LODs) for the two bacteria in foods were 2.05×10(3) cfu g(-1) and 1.04×10(3) cfu ml(-1), respectively. This immunosensor required only a bare electrode to measure the impedance changes, and no surficial modification on the electrode was needed. It was low-cost, reproducible, easy-to-operate, and easy-to-preserve. All these merits demonstrated this immunosensor has great potential for the rapid and on-site detection of pathogenic bacteria in foods.
authors.affiliation: Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Meng
authors.initials: M
authors.lastname: Xu
authors.affiliation: Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA.
authors.forename: Ronghui
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Biological and Agricultural Engineering, University of Arkansas, Fayetteville, AR 72701, USA; Center of Excellence for Poultry Science, University of Arkansas, Fayetteville, AR 72701, USA. Electronic address: yanbinli@uark.edu.
authors.forename: Yanbin
authors.initials: Y
authors.lastname: Li
mesh_terms: D000818:Animals; D002417:Cattle; D002645:Chickens / Q000382:microbiology; D015169:Colony Count, Microbial / Q000379:methods; D055664:Electrochemical Techniques / Q000379:methods*; D019453:Escherichia coli O157 / Q000302:isolation & purification*; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008839:Microelectrodes; D000069466:Red Meat / Q000382:microbiology; D012486:Salmonella typhimurium / Q000302:isolation & purification*; D013997:Time Factors
keywords: Electrochemical immunosensor; Escherichia coli O157:H7; Magnetic separation; Rapid detection; Salmonella Typhimurium; Screen-printed interdigitated microelectrode