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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 troubleshootingand 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.
List of contents
Traditional Methods of Analysis for Listeria monocytogenes.- MALDI-ToF MS: A Rapid Methodology for Identifying and Subtyping Listeria monocytogenes.- Sample Preparation for qPCR Detection of Listeria from Food.- qPCR Validation on the Basis of the Listeria monocytogenes prfA Assay.- Serotype Assignment by Sero-Agglutination, ELISA, and PCR.- Pulsed-Field Gel Electrophoresis (PFGE) Analysis of Listeria monocytogenes.- Multi-Locus Sequence Typing (MLST) and Whole Genome Sequencing (WGS) of Listeria monocytogenes and Listeria innocua.- High-Throughput Characterization of Listeria monocytogenes Using the OmniLog Phenotypic Microarray.- High-Throughput Screening of Biofilm Formation of Listeria monocytogenes on Stainless Steel Coupons Using a 96-Well Plate Format.- Confocal Laser Microscopy Analysis of Listeria monocytogenes Biofilms and Spatially Organized Communities.- Extraction and Preparation of Listeria monocytogenes Subproteomes for Mass Spectrometry Analysis.- Extraction and Analysis of Plasmid DNA from Listeria monocytogenes.- Generation of Nonpolar Deletion Mutants in Listeria monocytogenes Using the "SOEing" Method.- Mutant Construction and Integration Vector-Mediated Genetic Complementation in Listeria monocytogenes.- Internalization Assays for Listeria monocytogenes.- Microscopy of Intracellular Listeria monocytogenes in Epithelial Cells.- Control of Listeria monocytogenes Biofilms in a Simulated Food Processing Environment.- Sampling the Food Processing Environment: Taking up the Cudgel for Preventive Quality Management in Food Processing (FP).- Isolation and Evaluation of Anti-Listeria Lactococcus lactis from Vegetal Sources.
