Rapid Detection and Characterization of Foodborne Pathogens by Molecular Techniques: 1st Edition (Hardback) book cover

Rapid Detection and Characterization of Foodborne Pathogens by Molecular Techniques

1st Edition

By Robert E. Levin

CRC Press

608 pages | 25 B/W Illus.

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Hardback: 9781420092424
pub: 2009-10-26
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pub: 2009-10-26
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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.

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


Characteristics of the Genus

Virulence Factors

PCR Detection of Shigellae


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




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

Subject Categories

BISAC Subject Codes/Headings:
SCIENCE / Life Sciences / Biology / Microbiology