A microfluidic droplet digital PCR for simultaneous detection of pathogenic Escherichia coli O157 and Listeria monocytogenes.

摘要: Sensitive and rapid identification of pathogenic bacterial is extremely important due to the serious threat pathogens human health. In this study, we demonstrate simultaneous sensitive detection Escherichia coli O157 Listeria monocytogenes using a novel duplex droplet digital PCR (ddPCR) platform. The ddPCR platform, which uses mineral oil-saturated polydimethylsiloxane (OSP) chip overcome problem evaporation, integrates functions generation, on-chip amplification end-point fluorescence readout. Simultaneous two kinds achieved by design differentially labeled TaqMan-MGB fluorescent probes. Compared with quantitative real-time approach, OSP chip-based platform exhibits high sensitivity, at level single molecule resolution without significant cross-assay interference. Moreover, applicability proposed method also evaluated in artificially contaminated drinking water sample, displays low limit down 10 CFU/mL for both within 2 h.