Ultrasensitive quantification of pathogens in milliliters of beverage by filtration-based digital LAMP

Abstract

The quantitative detection of pathogens in milliliters of beverage sample requires complex preprocessing. To achieve rapid and ultrasensitive quantification of pathogens in large volume food sample, we developed a filtration-based interfacial digital LAMP (idLAMP) system, which consists of a nanoporous membrane for filtration and nanoporous hydrogel for digital amplification. Digital counting of single bacteria at the membrane surface under nanoconfinement could be achieved. The idLAMP successfully accomplished the quantitative detection of Escherichia coli in 100 mL water samples within 30 min, with wide dynamic range from 0.09 to 900 cells/mL. This technique could also be well applied to the quantification of Escherichia coli and Salmonella typhi in real beverage samples (juice, tea drinks, carbonated drinks and alcoholic drinks) without tedious sample pretreatments. With facile operation, higher specificity and sensitivity and better end-point analysis capabilities, the system has great potential in quantitative counting of single bacteria in large-volume food samples.

Keywords: Digital LAMP; Filtration; Hydrogel; Large volume; Nanoconfinement; Pathogen detection.