Group A Streptococcus, 1st ed. 2020
Methods and Protocols
Methods in Molecular Biology Series, Vol. 2136

This volume details Group A Streptococcus (GAS) research and provides the reader with an extensive collection of research protocols within this important field. Chapters guide readers through standard genetic protocols such as whole genome sequencing, transcriptome analyses, proteome analysis, application of GAS-derived molecular tools, and methods that are crucial for the evaluation of novel GAS vaccines and GAS virulence factors, including bactericidal assays and animal infection models. Written in the highly successful Methods in Molecular Biology series format, 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 and cutting-edge, Group A Streptococcus: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Part I: Genotyping and Genetic Manipulation of Group A Streptococcus

1. Detection of Streptococcus pyogenes Virulence Factors

Aleksandra Kozińska and Izabela Sitkiewicz

2. Closed-Tube Multiplex Real-Time PCR for the Detection of Group A Streptococcal Superantigens

Nour Zahi Gammoh andLothar Rink

3. The emm-cluster Typing System

4. Protocols for Tn-seq Analyses in The Group A Streptococcus

Yoann Le Breton, Ashton T. Belew, and Kevin S. McIver

5. Genetic Manipulation of Group A Streptococcus – Gene Deletion by Allelic Replacement

Timothy C. Barnett, Jessica N. Daw, Mark J. Walker, and Stephan Brouwer

6. Generation of Bioluminescent Group A Streptococcus for Biophotonic Imaging

Jacelyn M. S. Loh, Kar Yan Soh, and Thomas Proft

Part II: Application of Group A Streptococcus Omics

7. Whole Genome Sequence Analysis and Population Genomics of Group A streptococci

Jake A Lacey, Taylah B James, Steven YC Tong, and Mark R Davies

8. Group A Streptococcus Transcriptome Analysis

Misú Sansonand Anthony R. Flores

9. Protocol for Proteome Analysis of Group A Streptococcus

Laura Wilk

10. Investigation of Group A Streptococcal Interactions with Host Glycan Structures using High Throughput Techniques: Glycan Microarray Analysis using Recombinant Protein and Whole Cells

Anuk Indraratna, David De Oliveira, Lauren E Hartley –Tassell, Christopher J Day, Mark J Walker, Michael P Jennings,  and Martina L Sanderson-Smith

Part III: Assays to Investigate Group A Streptococcus Virulence Factors And Virulence Mechanisms

11. Using Lactococcus lactis as Surrogate Organism to Study Group A Streptococcus Surface Proteins

Catherine (Jia-Yun) Tsai, Jacelyn M.S. Loh, Sam Blanchett, and Thomas Proft

12. Expression and Purification of Collagen-like Proteins of Group A Streptococcus

Slawomir Lukomski and Dudley H. McNitt

13. Detection of Fibronectin-binding Proteins of Streptococcus pyogenes using Ligand Blot Analysis

Masanobu Nakata and Shigetada Kawabata

14. Morphology and Ultrastructure of Group A Streptococcus  Biofilms

Tatyana A. Danilova, T.A. Smirnova, G.A. Danilina, A.A. Adzhieva, N.V. Shevlyagina, and V.G. Zhukhovitsky

Angelica Riestra and J. Andrés Valderrama

16. Identification of Group A Streptococcus-containing autophagosome-like vacuoles 

Takashi Nozawa and Ichiro Nakagawa

17. Flow Cytometry-based Assays to Quantify Complement Deposition and Neutrophil Uptake of Group a Streptococcus

Nicola N.Lynskey

18. Live Cell Microscopy and Flow Cytometry to study Streptolysin S Mediated Erythrocyte Hemolysis 

Dustin L. Higashi, Charles R. Tessier, and Shaun W. Lee

19. Isolation of Monoclonal Antibodies to Group A Streptococcus Antigens Using Phage Display

Jeremy M. Raynes, Mei Lin Tay, Sok H. By, John D. Steemson, and Nicole J. Moreland

20. Assays to Analyse Adhesion of Group A Streptococcus to Host Cells

Adrina H. J. Khemlani, Thomas Proft, and Jacelyn M.S. Loh

21. The use of Galleria mellonella (wax moth) as an Infection Model for Group A Streptococcus

Catherine (Jia-Yun) Tsai, Jacelyn M.S. Loh, and Thomas Proft

22. A Superficial Skin Scarification Method in Mice to Mimic Streptococcus pyogenes Skin Infection in Humans

Manisha Pandey and Michael F Good

23. A Mouse Nasopharyngeal Colonisation Model for Group A Streptococcus

Adrina H. J. Khemlani, Thomas Proft, and Jacelyn M. S. Loh

24. Humanised Plasminogen Mouse Model to Study Group A Streptococcus Invasive Disease

Tania Rivera-Hernandez and Mark J. Walker

25. Lacefield Whole Blood Killing Assay to Evaluate Vaccine Efficacy

Mark Reglinski

26. An Opsonophagocytic Killing Assay for the Evaluation of Group A Streptococcus Vaccine Antisera

Reuben McGregor , Scott Jones , Jeremy Raynes , David Goldblatt , and Nicole J. Moreland

Part V: Group A Streptococcus Therapies

27. Validation of Suitable Carrier Molecules and Target Genes for antisense Therapy using Peptide-coupled Peptide Nucleic Acids (PNAs) in Streptococci

Gina Barkowsky, Bernd Kreikemeyer, and Nadja Patenge

28. Interfering with the Folding of Group A Streptococcal pili Proteins

Fernanda Contreras and Jaime Andrés Rivas-Pardo

Part VI: Application of Group A Streptococcus-derived Methods

29. Use of Streptolysin O (SLO) to Study the Function of Lipid Rafts

Heather E. Miller

30. Engineering of Group A Streptococcus Isopeptide Bonds into Immunoglobulin-like Protein Domains

Paul G. Youngand Edward N. Baker

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts