Listeria Monocytogenes Methods and Protocols

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. 

Part I: Detection, Quantification, and Confirmation

 

1. Traditional Methods of Analysis for Listeria monocytogenes

            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 monocytogenes prfA 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 Listeria monocytogenes

            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 Listeria monocytogenes 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-Listeria Lactococcus lactis from Vegetal Sources

            Van Thi Thuy Ho, Anran Dong, Raquel Lo, and Mark S. Turner