Chlamydia trachomatis: Methods and Protocols

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. Authoritative, practical, and relevant, Chlamydia trachomatis: Methods and Protocols serves as an ideal reference for scientists searching for a better understanding of the pathogen, allowing for the development of improved treatment regimens and the discovery of new drugs. Front Matter ....Pages i-xi Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Using Multiplex Strand Invasion Based Amplification (mSIBA) (Kevin E. Eboigbodin, Mark Hoser)....Pages 1-9 Point-of-Care Chlamydia trachomatis Detection Using Loop-Mediated Isothermal Amplification and Hydroxynaphthol Blue (Naraporn Somboonna, Ilada Choopara)....Pages 11-17 Application of Loop-Mediated Isothermal Amplification Assay for the Detection of Chlamydia trachomatis and Neisseria gonorrhoeae (Kevin E. Eboigbodin)....Pages 19-25 Simultaneous Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Using Real-Time Multiplex qPCR Assay (Kevin E. Eboigbodin)....Pages 27-32 Antimicrobial Resistance Screening in Chlamydia trachomatis by Optimized McCoy Cell Culture System and Direct qPCR-Based Monitoring of Chlamydial Growth (Tomislav Meštrović, Dezső P. Virok, Sunčanica Ljubin-Sternak, Tímea Raffai, Katalin Burián, Jasmina Vraneš)....Pages 33-43 Whole-Genome Sequencing of Chlamydia trachomatis Directly from Human Samples (Amanda Claire Brown, Mette T. Christiansen)....Pages 45-67 Multilocus Sequence Typing (MLST) of Chlamydiales (Martina Jelocnik, Adam Polkinghorne, Yvonne Pannekoek)....Pages 69-86 Identification and Discrimination of Chlamydia trachomatis Ocular and Urogenital Strains and Major Phylogenetic Lineages by CtGEM Typing, A Double-Locus Genotyping Method (Deborah C. Holt, Patiyan Andersson, Cameron Buckley, David M. Whiley, Philip M. Giffard)....Pages 87-122 Dual RNA-Seq of Chlamydia and Host Cells (James W. Marsh, Regan J. Hayward, Amol Shetty, Anup Mahurkar, Michael S. Humphrys, Garry S. A. Myers)....Pages 123-135 Isolation and Propagation of Single Inclusion-Derived Chlamydia Using Laser Microdissection (Oleg V. Podgorny, Nadezhda F. Polina, Vassili N. Lazarev)....Pages 137-150 Genetic Manipulation of Chlamydia trachomatis: Chromosomal Deletions (Katerina Wolf, Mostafa Rahnama, Kenneth A. Fields)....Pages 151-164 Mutagenesis of Chlamydia trachomatis Using TargeTron (Mary M. Weber, Robert Faris)....Pages 165-184 Forward and Reverse Genetic Analysis of Chlamydia (Mateusz Kędzior, Robert J. Bastidas)....Pages 185-204 A Coinfection Model to Evaluate Chlamydia Inc Protein Interactions (Rachel Ende, Isabelle Derré)....Pages 205-218 Rodent Infections for Chlamydia spp. (Charles W. Armitage, Alison J. Carey, Kenneth W. Beagley)....Pages 219-236 Enumeration of Viable Chlamydia from Infected Animals Using Immunofluorescent Microscopy (Steven Liang, James B. Mahony)....Pages 237-244 Proximity Labeling of the Chlamydia trachomatis Inclusion Membrane (Macy G. Olson, Lisa M. Jorgenson, Ray E. Widner, Elizabeth A. Rucks)....Pages 245-278 High-Throughput Screening for Novel Inhibitors of Intracellular Pathogens, Including Chlamydia trachomatis (Amanda Claire Brown, Nicole L. Kushner)....Pages 279-286 Back Matter ....Pages 287-289