Journal
BIOSENSORS & BIOELECTRONICS
Volume 74, Issue -, Pages 770-777Publisher
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2015.07.016
Keywords
Droplet digital PCR; Pathogenic bacterial; Simultaneous detection; Escherichia coli O157; Listeria monocytogenes
Categories
Funding
- National Science Foundation of China [21405167, 61271162, 81472751]
- National High Technology Research and Development Program (863Program) of China [2012BAK08B05]
- Science and Technology Commission of Shanghai Municipality [15ZR1447000, 11JC1414400]
- Jiangsu Key Laboratory of Biomaterials and Devices [LBMD201401]
- Open Research Fund of State Key Laboratory of Bioelectronics, Southeast University, Nanjing
Ask authors/readers for more resources
Sensitive and rapid identification of pathogenic bacterial is extremely important due to the serious threat of pathogens to human health. In this study, we demonstrate the simultaneous and sensitive detection of pathogenic Escherichia coil O157 and Listeria monocytogenes using a novel duplex droplet digital PCR (ddPCR) platform. The ddPCR platform, which uses a mineral oil-saturated polydimethylsiloxane (OSP) chip to overcome the problem of droplet evaporation, integrates the functions of droplet generation, on-chip amplification and end-point fluorescence readout. Simultaneous detection of two kinds of bacterial is achieved by the design of differentially labeled TaqMan-MGB fluorescent probes. Compared with a quantitative real-time PCR approach, the OSP chip-based duplex ddPCR platform exhibits high sensitivity, which is at the level of single molecule resolution without significant cross-assay interference. Moreover, the applicability of the proposed method is also evaluated in artificially contaminated drinking water sample, which displays a low detection limit down to 10 CFU/mL for both pathogenic bacterial within 2 h. (C) 2015 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available