Main 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.

Contents:

1. Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Using Multiplex Strand Invasion-Based Amplification (mSIBA)

Kevin E. Eboigbodin and Mark Hoser

2. Point of Care Chlamydia trachomatis Detection Using Loop-Mediated Isothermal Amplification and Hydroxynaphthol Blue

Naraporn Somboonna and Ilada Choopara

3. Application of Loop-Mediated Isothermal Amplification Assay for the Detection of Chlamydia trachomatis and Neisseria gonorrhoeae

Kevin E. Eboigbodin

4. Simultaneous Detection of Chlamydia trachomatis and Neisseria gonorrhoeae Using Real-Time Multiplex qPCR Assay

Kevin E. Eboigbodin

5. Antimicrobial Resistance Screening in Chlamydia trachomatis by Optimized McCoy Cell Culture System and Direct qPCR-Based Monitoring of Chlamydial Growth

Tomislav Mestrovic, Dezso P. Virok, Suncanica Ljubin-Sternak, Timea Raffai, Katalin Burian, and Jasmina Vranes

6. Whole Genome Sequencing of Chlamydia trachomatis Directly from Human Samples

Amanda Claire Brown and Mette T. Christiansen

7. Multi Locus Sequence Typing (MLST) of Chlamydiales

Martina Jelocnik, Adam Polkinghorne, and Yvonne Pannekoek

8. 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, and Phillip M. Giffard

9. Dual RNA-Seq of Chlamydia and Host Cells

James W. Marsh, Regan Hayward, Amol Shetty, Anup Mahurkar, Michael S. Humphrys, and Garry S.A. Myers

10. Isolation and Propagation of Single Inclusion-Derived Chlamydia Using Laser Microdissection

Oleg V. Podgorny, Nadezhda F. Polina, and Vassili N. Lazarev

11. Genetic Manipulation of Chlamydia trachomatis: Chromosomal Deletions

Katerina Wolf, Mostafa Rahnama, and Kenneth A. Fields

12. Mutagenesis of Chlamydia trachomatis Using TargeTron

Mary M. Weber and Robert Faris

13. Forward and Reverse Genetic Analysis of Chlamydia

Mateusz Kedzior and Robert J. Bastidas

14. A Co-Infection Model to Evaluate Chlamydia Inc Protein Interactions

Rachel Ende and Isabelle Derre

15. Rodent Infections for Chlamydia spp.

Charles W. Armitage, Alison J. Carey, and Kenneth W. Beagley

16. Enumeration of Viable Chlamydia from Infected Animals Using Immunofluorescent Microscopy

Steven Liang and James B. Mahony

17. Proximity Labeling of the Chlamydia trachomatis Inclusion Membrane

Macy G. Olson, Lisa M. Jorgenson, Ray E. Widner, and Elizabeth A. Rucks

18. High-Throughput Screening for Novel Inhibitors of Intracellular Pathogens, including Chlamydia trachomatis

Amanda Claire Brown and Nicole L. Kushner