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KMID : 0380620160480050454
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Korean Journal of Food Science and Technology
2016 Volume.48 No. 5 p.454 ~ p.460
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Detection and quantitation of Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli O157:H7 by droplet digital PCR
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Kim Jin-Hee
Yoon Jin-Sun
Lee Da-Young
Kim Dong-ho
Oh Se-Wook
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Abstract
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In this study, we investigated the possibility of Droplet digital PCR (ddPCR) for detection of foodborne pathogens. ddPCR combines partitioning of PCR reactions into several thousands or millions of individual droplets in a water-oil emulsion, and counting of positive PCR reaction using flow cytometry. Four species of foodborne pathogens, Bacillus cereus, Staphylococcus aureus, Salmonella Typhimurium and Escherichia coli O157:H7, were used to quantify the target sequence with each of the designed primers and double stranded DNA-binding Evagreen dye. All tested foodborne pathogens showed a detection limit ranging from 100 fg/μL to 10 ng/μL. It was concluded that ddPCR could be used to detect very low concentrations of foodborne pathogens from complex food matrices. For multi-detection of target pathogens, we also tested the samples using multiplex ddPCR and obtained successful results.
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KEYWORD
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droplet digital PCR, water-oil emulsion, pathogens, flow cytometry, detection
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