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The swab sampling method combined with detection approaches is a common approach used to monitor pathogenic microbes in food facilities and to determine the adequateness of regular cleaning and sanitation processes. Thus, the efficiency of a swab material for both the capture and release of bacteria has a significant impact on the detection of pathogens. A novel functionalized cotton swab was developed based on cationic functionalization of the cotton swab for improving the sampling of bacteria from surfaces. The cationic modified swab was fabricated using UV-induced grafting of cationic monomer (METAC) onto a regular cotton swab. The prepared cationic cotton swabs displayed a persistent positive charge regardless of the pH conditions. The cationic swabs demonstrated 150–200% enhancement in the swab performance for Gram-positive and Gram-negative bacteria under any sampling conditions compared to the regular cotton swab as well as a significant enhancement in the bacteria sampling at the low bacterial concentrations. Furthermore, the cationic cotton swabs showed significant improvement at least 4 folds in the sampling performance of the bacterial biofilm from the stainless-steel surfaces compared to the cotton swabs. These results illustrate the role of modified swabs in enhancing the sampling of bacteria from contact surfaces and their potential impact on improved monitoring of microbial contamination and verification of surface sanitization.

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