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Title: Light-scattering sensor for real-time identification of Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae colonies on solid agar plate
Authors: Karleigh Huff
Amornrat Aroonnual
Amy E.Fleishman Littlejohn
Bartek Rajwa
Euiwon Bae
Padmapriya P. Banada
Valery Patsekin
E. Daniel Hirleman
J. Paul Robinson
Gary P. Richards
Arun K. Bhunia
Purdue University
USDA Agricultural Research Service, Washington DC
Virginia Polytechnic Institute and State University
Mahidol University
Rutgers New Jersey Medical School
UC Merced
Keywords: Biochemistry, Genetics and Molecular Biology;Chemical Engineering;Immunology and Microbiology
Issue Date: 1-Sep-2012
Citation: Microbial Biotechnology. Vol.5, No.5 (2012), 607-620
Abstract: The three most common pathogenic species of Vibrio, Vibrio cholerae, Vibrio parahaemolyticus and Vibrio vulnificus, are of major concerns due to increased incidence of water- and seafood-related outbreaks and illness worldwide. Current methods are lengthy and require biochemical and molecular confirmation. A novel label-free forward light-scattering sensor was developed to detect and identify colonies of these three pathogens in real time in the presence of other vibrios in food or water samples. Vibrio colonies grown on agar plates were illuminated by a 635nm laser beam and scatter-image signatures were acquired using a CCD (charge-coupled device) camera in an automated BARDOT (BActerial Rapid Detection using Optical light-scattering Technology) system. Although a limited number of Vibrio species was tested, each produced a unique light-scattering signature that is consistent from colony to colony. Subsequently a pattern recognition system analysing the collected light-scatter information provided classification in 1-2min with an accuracy of 99%. The light-scattering signatures were unaffected by subjecting the bacteria to physiological stressors: osmotic imbalance, acid, heat and recovery from a viable but non-culturable state. Furthermore, employing a standard sample enrichment in alkaline peptone water for 6h followed by plating on selective thiosulphate citrate bile salts sucrose agar at 30°C for ~12h, the light-scattering sensor successfully detected V.cholerae, V.parahaemolyticus and V.vulnificus present in oyster or water samples in 18h even in the presence of other vibrios or other bacteria, indicating the suitability of the sensor as a powerful screening tool for pathogens on agar plates. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.
ISSN: 17517915
Appears in Collections:Scopus 2011-2015

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