Rapid detection of respiring Escherichia coli O157:H7 in apple juice, milk, and ground beef by flow cytometry

Cytometry. Part a : the Journal of the International Society for Analytical Cytology
N YamaguchiMasao Nasu

Abstract

Rapid and simple methods to detect viable pathogenic microbes in foods and drinks are required. Flow cytometry was used for the rapid detection of respiring Escherichia coli O157:H7 cells in apple juice, milk, and ground beef. CTC (5-cyano-2,3-ditolyl tetrazolium chloride) was used to estimate the respiratory activity of bacteria. Fluorescein isothiocyanate (FITC)-labeled anti-E. coli O157:H7 direct antibody (FA) was used for the specific detection of target cells. Food samples were inoculated with starved E. coli O157:H7 and E. coli K-12 cells, and analyzed by both fluorescent microscopy and flow cytometry after double staining with FA and CTC. Respiring E. coli O157:H7 cells in food samples showed strong fluorescence of both FA (green) and CTC (red); thus, they could be clearly and specifically distinguished from respiring E. coli K-12 or inactive cells. A good correlation was achieved in flow cytometric analysis between the numbers of inoculated viable E. coli O157:H7 and those detected in milk and apple juice. The detection threshold for this flow cytometry for E. coli O157:H7 in milk, apple juice, and ground beef was 10(3) cells/ml (milk and apple juice) or 10(3) cells/g (ground beef) of sample when the total bacterial num...Continue Reading

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Dec 20, 2008·Analytical and Bioanalytical Chemistry·F Ceyda DudakWilliam R Heineman
Apr 20, 2010·Analytical and Bioanalytical Chemistry·Erhan TemurNihal Aydogan
May 25, 2010·Journal of Industrial Microbiology & Biotechnology·Robert Scott DonofrioSusan T Bagley
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Related Concepts

5-cyano-2,3-ditolyltetrazolium chloride
Teas, Medicinal
Bos indicus
Alkalescens-Dispar Group
Flow Cytometry
Food Contamination
Food Microbiology
Meat
Staining and Labeling
Tetrazolium Salts

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