A guanidinium-rich polymer as a new universal bioreceptor for multiplex detection of bacteria from environmental samples

Protamine, a guanidinium rich polymer, is proposed as a universal bioreceptor for bacteria, towards rapid and handheld bacteria detection from complex environmental water samples without the need for specific antibodies or primers. Escherichia coli K12, Salmonella Typhimurium, and Staphylococcus aureus (MSSA) were assayed, representing gram-negative, gram-positive, rod- and round-shaped bacteria. Samples and the protamine conjugated fluorescent particles were sequentially loaded to the paper microfluidic chips and flowed through the channels spontaneously via capillary action. The particles were aggregated via protamine-bacteria membrane interactions and unbound particles were rinsed via capillary action. A low-cost smartphone fluorescence microscope was designed, fabricated, and imaged the paper channels. A unique image processing algorithm isolated only the aggregated particles to detect all three bacteria (p < 0.05) with a detection limit of 10¹−10² CFU/mL. Protamine did not induce any particle aggregation with a model protein, algae, and virus. Successful bacteria detection was also demonstrated with environmental field water samples. Total assay time was < 10 min with neither extraction nor enrichment steps. In summary, a guanidinium-rich polymer showed a promise as a universal bioreceptor for bacteria and can be used on a paper microfluidic chip and smartphone quantification towards rapid and handheld detection.