Description
This book explores cutting-edge methods to work with the notoriously difficult, but highly prevalent, obligate intracellular pathogen, Chlamydia trachomatis. These include techniques to identify Chlamydia trachomatis in patient samples, ranging from simple point-of-care tests to whole genome sequencing; methods for propagation of strains in both cell culture and animal models; techniques to manipulate Chlamydia trachomatis in molecular genetic methodologies; a high-throughput screening method for testing new potential drugs against intracellular bacteria; a screen for antibiotic resistance; methods for labeling and enumeration; and descriptions of genotyping technologies, as well as dual RNA-Seq transcriptional profiling. 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.
Table of Contents
Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Using Multiplex Strand Invasion-Based Amplification (mSIBA).- Point of Care Chlamydia trachomatis Detection Using Loop-Mediated Isothermal Amplification and Hydroxynaphthol Blue.- Application of Loop-Mediated Isothermal Amplification Assay for the Detection of Chlamydia trachomatis and Neisseria gonorrhoeae.- Simultaneous Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Using Real-Time Multiplex qPCR Assay.- Antimicrobial Resistance Screening in Chlamydia trachomatis by Optimized McCoy Cell Culture System and Direct qPCR-Based Monitoring of Chlamydial Growth.- Whole Genome Sequencing of Chlamydia trachomatis Directly from Human Samples.- Multi Locus Sequence Typing (MLST) of Chlamydiales.- Identification and Discrimination of Chlamydia trachomatis Ocular and Urogenital Strains and Major Phylogenetic Lineages by CtGEM Typing, a Double Locus Genotyping Method.- Dual RNA-Seq of Chlamydia and Host Cells.- Isolation and Propagation of Single Inclusion-Derived Chlamydia Using Laser Microdissection.- Genetic Manipulation of Chlamydia trachomatis: Chromosomal Deletions.- Mutagenesis of Chlamydia trachomatis Using TargeTron.- Forward and Reverse Genetic Analysis of Chlamydia.- A Co-Infection Model to Evaluate Chlamydia Inc Protein Interactions.- Rodent Infections for Chlamydia spp..- Enumeration of Viable Chlamydia from Infected Animals Using Immunofluorescent Microscopy.- Proximity Labeling of the Chlamydia trachomatis Inclusion Membrane.- High-Throughput Screening for Novel Inhibitors of Intracellular Pathogens, including Chlamydia trachomatis.
