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More About This Textbook
Overview
Decades of development of the polymerase chain reaction (PCR) have yielded a significant array of associated techniques that make it possible to rapidly detect low numbers of all known pathogenic microorganisms without the traditional, more taxing methods of cultivation and phenotypic characterization. Written by one of the most prolific and respected researchers in food safety, Rapid Detection and Characterization of Foodborne Pathogens by Molecular Techniques describes the application of molecular techniques for the detection and discrimination of major infectious bacteria associated with foods.
The book puts a particular focus on genes associated with pathogenicity used in PCR, including real-time PCR for specific detection of pathogenic bacteria and the inherent limitations of such methodology with certain pathogens. It also emphasizes methods for extracting microorganisms from complex food matrices and DNA purification techniques. The coverage begins with a highly comprehensive review of real time PCR, complete with theoretical and operational concepts. Each chapter deals with a specific organism and the techniques applied to that organism. The text includes references on the use of PCR primers and DNA probes, the DNA sequence of each being listed at the end of each chapter to create a complete compendium.
This is not a "recipe book", but rather a resource with sufficiently detailed information that allows readers to fully comprehend the methodology described and the significance of the results. Copiously illustrated with figures, tables, charts, and graphs, this is a detailed presentation of the major, contemporary studies involving the molecular detection, quantification, and subspecies differentiation of each organism. With objective assessments of the molecular techniques, their advantages, and limitations, the book allows investigators to readily identify the precise molecular technique and application most suitable for their research.
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Table of Contents
Molecular Techniques for Detecting, Quantifying, and Subspecies Typing of Foodborne Pathogenic Bacteria
The Polymerase Chain Reaction (PCR)
Real-Time PCR (Rti-PCR)
Nested PCR
Loop-Mediated Isothermal Amplification (LAMP) Assay
Pulsed-Field Gel Electrophoresis (PFGE)
Random Amplification of Polymorphic DNA (RAPD)
Multilocus Sequence Typing (MLST)
Resriction Fragment Length Polymorphism (RFLP) and PCR-RFLP
Amplified Fragment Length Polymorphism (AFLP)
Escherichia coli O157:H7
Characteristics of the Organism
Virulence Factors
Phenotypic Variation of E. coli O157:H7
PCR Assays Involving Molecular Probes and Real-Time PCR (Rti-PCR)
Loop-Mediated Isothermal Amplification (LAMP) of DNA
Immunomagnetic Separation and PCR
Restriction Fragment Length Polymorphism
Subspecies Typing of E. coli O157:H7 Isolates
Shigella
Characteristics of the Genus
Virulence Factors
PCR Detection of Shigellae
Salmonella
Characteristics of the Genus
Molecular Techniques
Vibrio vulnificus
Characteristics of the Organism
Virulence Factors
Enrichment and Isolation Media
Identification of V. vulnificus Isolates
Typing of V. vulnificus Isolates below the Species Level
Molecular Methods for Detection and Typing
Vibrio parahaemolyticus
Characteristics of the Organism
Clinical Symptoms Due to Infections by V. parahaemolyticus
Ecology of V. parahaemolyticus
Phenotypic Characteristics of V. parahaemolyticus
Sensitivity of V. parahaemolyticus to Low Temperatures
Isolation and Cultivation of V. parahaemolticus
Preservation of V. parahaemolyticus Isolates
Bacteriophage for V. parahaemolyticus
Use of the Polymerase Chain Reaction (PCR) for Detection of V. parahaemolyticus
Molecular Typing of V. parahaemolyticus Isolates below the Species Level
The O3:K6 pandemic clone
Vibrio cholera
Characteristics of the Organism
Factors Associated with the Virulence of V. cholerae
PCR Detection, Identification, and Characterization of V. cholerae
Molecular Typing of V. cholerae Isolates
Acquisition of Antibiotic Resistance
Biotype Conversion by Phage
Aeromonas hydrophila
Characteristics of the Organism
PCR Detection
Distribution of Toxin Genes
Genotyping Using Hemolysin and Aerolysin Genes
Restriction Fragment Length Polymorphism (RFLP)
Plesiomonas shigelloides
Characteristics of the Organism
Nomenclature and Taxonomy
Physiological and Biochemical Characteristics
Ecological Distribution
Toxins and Invasive Factors
β-Hemolysis
Isolation
Serology
Epidemiology and Outbreaks
Application of the Polymerase Chain Reaction (PCR)
Campylobacter jejuni
Characteristics of the Organism
Ecological Distribution of Campylobacters
Virulence Factors
Isolation of Campylobacters from Foods
Serotyping of Campylobacters and Immunological Detection
Bacteriophage Typing of C. jejuni
Molecular Methods of Detectng and Typing Campylobacters
The Viable but Nonculturable (VBNC) State of Campylobacters
The Coccoid form of C. jejuni
Immunomagnetic Capture of C. jejuni
Staphylococcus aureus
Characteristics of the Organism
Molecular Techniques for Detection and Identification of S. aureus
The Role of S. aureus Bacteriophage (Phage) in Pathogenesis
Listeria monocytogenes
Characteristics of the Organism
Clostridium botulinum
Characteristics of the Organism
Relationship between Botulism and Seafood
Infant Botulism
Molecular Techniques Applied to C. botulinum
Clostridium perfringens
Characteristics of the Organism
Molecular Methods for Detection of C. perfringens
Molecular Typing of Isolates
Bacillus cereus
Characteristics of the Organism
Selective Isolation of B. cereus
The Emetic Toxin Cereulide
Enterotoxins of B. cereus
Molecular Methods for Detection and Confirmation of B. cereus
Molecular Typing of B. cereus Isolates