1. Multipurpose assessment for the quantification of Vibrio spp. and total bacteria in fish and seawater using multiplex real-time polymerase chain reaction.
- Author
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Kim JY and Lee JL
- Subjects
- Animals, Aquaculture, Atlantic Ocean, Colony Count, Microbial, Conserved Sequence, DNA, Bacterial metabolism, DNA, Ribosomal metabolism, Databases, Nucleic Acid, Delaware, Gram-Negative Bacteria classification, Gram-Negative Bacteria growth & development, Gram-Negative Bacteria metabolism, Gram-Positive Bacteria classification, Gram-Positive Bacteria growth & development, Gram-Positive Bacteria metabolism, Molecular Typing, Multiplex Polymerase Chain Reaction, RNA, Ribosomal, 16S metabolism, Real-Time Polymerase Chain Reaction, Seafood economics, Vibrio classification, Vibrio growth & development, Vibrio metabolism, Vibrio parahaemolyticus classification, Vibrio parahaemolyticus growth & development, Vibrio parahaemolyticus isolation & purification, Vibrio parahaemolyticus metabolism, Vibrio vulnificus classification, Vibrio vulnificus growth & development, Vibrio vulnificus isolation & purification, Vibrio vulnificus metabolism, Fishes microbiology, Food Inspection methods, Gram-Negative Bacteria isolation & purification, Gram-Positive Bacteria isolation & purification, Seafood microbiology, Seawater microbiology, Vibrio isolation & purification
- Abstract
Background: This study describes the first multiplex real-time polymerase chain reaction assay developed, as a multipurpose assessment, for the simultaneous quantification of total bacteria and three Vibrio spp. (V. parahaemolyticus, V. vulnificus and V. anguillarum) in fish and seawater. The consumption of raw finfish as sushi or sashimi has been increasing the chance of Vibrio outbreaks in consumers. Freshness and quality of fishery products also depend on the total bacterial populations present., Results: The detection sensitivity of the specific targets for the multiplex assay was 1 CFU mL⁻¹ in pure culture and seawater, and 10 CFU g⁻¹ in fish. While total bacterial counts by the multiplex assay were similar to those obtained by cultural methods, the levels of Vibrio detected by the multiplex assay were generally higher than by cultural methods of the same populations. Among the natural samples without Vibrio spp. inoculation, eight out of 10 seawater and three out of 20 fish samples were determined to contain Vibrio spp., Conclusion: Our data demonstrate that this multiplex assay could be useful for the rapid detection and quantification of Vibrio spp. and total bacteria as a multipurpose tool for surveillance of fish and water quality as well as diagnostic method., (© 2014 The Authors. Journal of the Science of Food and Agriculture published by JohnWiley & Sons Ltd on behalf of Society of Chemical Industry.)
- Published
- 2014
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