Listeria monocytogenes Methods and Protocols

Listeria monocytogenes is still a major threat to public health. A new book in the Methods in Molecular Biology series, Listeria monocytogenes: Methods and Protocols addresses its titular pathogen with protocols and methodologies used in research to gain a better understanding of Listeria at a molecular level. The topics covered include sampling in order to isolate Listeria, methods for their identification and characterization, methods for gene manipulation and finally, methods for control of the organism. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols and tips on troubleshooting and avoiding known pitfalls.

Vital and authoritative, Listeria monocytogenes: Methods and Protocols aims to contribute to the harmonization of the methods used in the field and will therefore benefit all those interested in Listeria research.

Part I: Detection, Quantification, and Confirmation

1. Sampling the Production/Processing Environment for Listeria

            Anca Ioana Nicolau and Andrei Sorin Bolocan

2. Traditional Methods for Isolation of Listeria monocytogenes

            Rui Magalhães, Cristina Mena, Vânia Ferreira, Gonçalo Almeida, Joana Silva, and Paula Teixeira

3. Confirmation of Isolates of Listeria by Conventional and Real-Time PCR

            David Rodríguez-Lázaro and Marta Hernández

Part II: Characterization and Typing

4. Serotype Assignment by Sero-Agglutination, ELISA, and PCR

            Lisa Gorski

5. Pulsed-Field Gel Electrophoresis (PFGE) Analysis of Listeria monocytogenes

            Marion Dalmasso and Kieran Jordan

6. Multi-Locus Sequence Typing (MLST) of Listeria monocytogenes

            Beatrix Stessl, Irene Rückerl, and Martin Wagner

7. Ribotyping and Automated Ribotyping of Listeria monocytogenes

            Mazin Matloob and Mansel Griffiths

8. Fluorescent Amplified Fragment Length Polymorphism (fAFLP) Analysis of Listeria monocytogenes

            Corinne Amar

9. High-Throughput Characterization of Listeria monocytogenes Using the OmniLog Phenotypic Microarray

            Edward M. Fox and Kieran Jordan

10. Analysis of Listeria monocytogenes Subproteomes

            Michel Hébraud

11. The Listeria Cell Wall and Associated Carbohydrate Polymers

            Marcel R. Eugster and Martin J. Loessner

12. Use of Bacteriophage Cell Wall Binding Proteins for Rapid Diagnostics of Listeria

            Mathias Schmelcher and Martin J. Loessner

13. Virulence Characterization of Listeria monocytogenes

            Swetha Reddy and Mark L. Lawrence

14. Internalization Assays for Listeria monocytogenes

            Andreas Kühbacher, Pascale Cossart, and Javier Pizarro-Cerdá

Part III: Strain Manipulation

15. Extraction and Analysis of Plasmid DNA from Listeria monocytogenes

            Aidan Casey and Olivia McAuliffe

16. Generation of Non-Polar Deletion Mutants in Listeria monocytogenes Using the “SOEing” Method

            Kathrin Rychli, Caitriona M. Guinane, Karen Daly, Colin Hill, and Paul D. Cotter

17. Mutant Construction and Integration Vector-Mediated Gene Complementation in Listeria monocytogenes

            Reha Onur Azizoglu, Driss Elhanafi, and Sophia Kathariou

18. Absolute and Relative Gene Expression in Listeria monocytogenes Using Real Time PCR

            Roberta Mazza and Rina Mazette

19. Genome Sequencing of Listeria monocytogenes

            Stephan Schmitz-Esser and Martin Wagner

20. Using Enhanced Green Fluorescent Protein (EGFP) Promoter Fusions to Study Gene Regulation at Single Cell and Population Levels

            Marta Utratna and Conor P. O’Byrne

Part IV: Control Methods

21. Control of Listeria monocytogenes in the Processing Environment by Understanding Biofilm Formation and Resistance to Sanitizers

            Stavros G. Manios and Panagiotis N. Skandamis

22. Vaccination Studies: Detection of a Listeria monocytogenes Specific T Cell Immune Response Using the ELISPOT Technique

            Mohammed Bahey-El-Din and Cormac G.M. Gahan

23. Sampling the Food Processing Environment: Taking up the Cudgel for Preventive Quality Management in Food Processing Environments

            Martin Wagner and Beatrix Stessl