Part I: Detection, Quantification, and Confirmation
1. Traditional Methods of Analysis for Listeriamonocytogenes
Rui Magalhães, Cristina Mena, Vânia Ferreira, Gonçalo Almeida, Joana Silva, and Paula Teixeira
2. MALDI-ToF MS: A Rapid Methodology for Identifying and Subtyping Listeria monocytogenes
Snehal R. Jadhav, Rohan M. Shah, and Enzo A. Palombo
3. Sample Preparation for qPCR Detection of Listeria from Food
Patrick Mester, Anna Kristina Witte, and Peter Rossmanith
4. qPCR Validation on the Basis of the Listeria monocytogenesprfA Assay
Anna Kristina Witte, Patrick Mester, and Peter Rossmanith
Part II: Subtyping Approaches
5. Serotype Assignment by Sero-Agglutination, ELISA, and PCR
Lisa Gorski
6. Pulsed-Field Gel Electrophoresis (PFGE) Analysis of Listeria monocytogenes
Karen Hunt and Kieran Jordan
7. Multi-Locus Sequence Typing (MLST) and Whole Genome Sequencing (WGS) of Listeria monocytogenes and Listeria innocua
Beatrix Stessl, Martin Wagner, and Werner Ruppitsch
Part III: Genotypic and Phenotypic Characterization
8. High-Throughput Characterization of Listeria monocytogenes Using the OmniLog Phenotypic Microarray
Laura Luque-Sastre, Kieran Jordan, Séamus Fanning, and Edward M. Fox
9. High-Throughput Screening of Biofilm Formation of Listeria monocytogenes on Stainless Steel Coupons Using a 96-Well Plate Format
Jessica A. Gray, P. Scott Chandry, John P. Bowman, and Edward M. Fox
10. Confocal Laser Microscopy Analysis of Listeria monocytogenes Biofilms and Spatially Organized Communities
Maud Darsonval, Marina Grégoire, Julien Deschamps, and Romain Briandet
11. Extraction and Preparation of Listeria monocytogenes Subproteomes for Mass Spectrometry Analysis
Tiago Santos and Michel Hébraud
Part IV: Strain Manipulation
12. Extraction and Analysis of Plasmid DNA from Listeriamonocytogenes
Aidan Casey and Olivia McAuliffe
13. Generation of Nonpolar Deletion Mutants in Listeria monocytogenes Using the “SOEing” Method
Kathrin Rychli, Eva Harter, Caitriona M. Guinane, Karen Daly, Colin Hill, and Paul D. Cotter
14. Mutant Construction and Integration Vector-Mediated Genetic Complementation in Listeria monocytogenes
Cameron Parsons, Reha Azizoglu, Driss Elhanafi, and Sophia Kathariou
Part V: Host-Pathogen Interactions
15. Internalization Assays for Listeria monocytogenes
Andreas Kühbacher, Pascale Cossart, and Javier Pizarro-Cerdá
16. Microscopy of Intracellular Listeriamonocytogenes in Epithelial Cells
Hélène Bierne, Mounia Kortebi, and Natalie Descoeudres
Part VI: Control Methods
17. Control of Listeria monocytogenes Biofilms in a Simulated Food Processing Environment
Sofia V. Poimenidou, Stavros G. Manios, and Panagiotis N. Skandamis
18. Sampling the Food Processing Environment: Taking up the Cudgel for Preventive Quality Management in Food Processing (FP)
Martin Wagner and Beatrix Stessl
19. Isolation and Evaluation of Anti-ListeriaLactococcus lactis from Vegetal Sources
Van Thi Thuy Ho, Anran Dong, Raquel Lo, and Mark S. Turner
This updated book explores a wide repertoire of tools and approaches that have been created, modified, and applied to the study of L. monocytogenes, forming the basis of our understanding of the bacterium today. Many of these key experimental techniques are gathered together herein. The volume presents aspects such as clinical disease and host-pathogen interactions, as well as the study of biofilms which present a significant challenge for control of the organism in the food processing environment. The topics covered in this edition also include sampling in order to isolate Listeria, methods for their identification and characterization, methods for gene manipulation, and methods for control of the organism. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, Listeria monocytogenes: Methods and Protocols, Second Edition aims to contribute toward the harmonization of methods used to study this important bacterium, and to be of particular interest to Listeria research both in relation to food association and control as well as clinical microbiology.