Microfluidic paper-based aptasensor devices for multiplexed detection of pathogenic bacteria

Foodborne pathogens are major public health concerns worldwide. Paper-based microfluidic devices are versatile, user friendly and low cost. We report a novel paper-based single input channel microfluidic device that can detect more than one whole-cell foodborne bacteria at the same time, and comes with quantitative reading via image analysis. This microfluidic paper-based multiplexed aptasensor simultaneously detects E. coli O157:H7 and S. Typhimurium. Custom designed particles provide colorimetric signal enhancement and false results prevention. Several aptamers were screened and the highest-affinity aptamers were optimized and employed for detection of these bacteria in solution, both in a buffer as well as pear juice. Image analysis was used to read and quantify the colorimetric signal and measure bacteria concentration, thus rendering this paper based microfluidic device quantitative. The colorimetric results show linearity over a wide concentration range (10² CFU/mL to 10⁸ CFU/mL) and a limit of detection (LOD) of 10³ CFU/mL and 10² CFU/mL for E. coli O157:H7 and S. Typhimurium, respectively. An insignificant change in colorimetric response for non-target bacteria indicates the aptasesnors are specific. The reported multiplexed colorimetric paper-based microfluidic devices is likely to perform well for on-site rapid screening of pathogenic bacteria in water and food products.