1. A highly sensitive 3base™ assay for detecting Streptococcus pyogenes in saliva during controlled human pharyngitis.
- Author
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Indraratna AD, Mytton S, Ricafrente A, Millar D, Gorman J, Azzopardi KI, Frost HR, Osowicki J, Steer AC, Skropeta D, and Sanderson-Smith ML
- Subjects
- Humans, Streptococcal Infections diagnosis, Streptococcal Infections microbiology, Adult, Real-Time Polymerase Chain Reaction methods, Streptococcus pyogenes isolation & purification, Saliva microbiology, Pharyngitis microbiology, Pharyngitis diagnosis
- Abstract
Streptococcus pyogenes (Group A Streptococcus; GAS) is a Gram-positive bacterium responsible for substantial human mortality and morbidity. Conventional diagnosis of GAS pharyngitis relies on throat swab culture, a low-throughput, slow, and relatively invasive 'gold standard'. While molecular approaches are becoming increasingly utilized, the potential of saliva as a diagnostic fluid for GAS infection remains largely unexplored. Here, we present a novel, high-throughput, sensitive, and robust speB qPCR assay that reliably detects GAS in saliva using innovative 3base™ technology (Genetic Signatures Limited, Sydney, Australia). The assay has been validated on baseline, acute, and convalescent saliva samples generated from the Controlled Human Infection for Vaccination Against Streptococcus (CHIVAS-M75) trial, in which healthy adult participants were challenged with emm75 GAS. In these well-defined samples, our high-throughput assay outperforms throat culture and conventional qPCR in saliva respectively, affirming the utility of the 3base™ platform, demonstrating the feasibility of saliva as a diagnostic biofluid, and paving the way for the development of novel non-invasive approaches for the detection of GAS and other oropharyngeal pathogens., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Martina Sanderson-Smith reports financial support was provided by National Health and Medical Research Council. Anuk Indraratna reports financial support, administrative support, and equipment, drugs, or supplies were provided by Genetic Signatures Limited. Anuk Indraratna reports financial support was provided by Australian Government Department of Education. Sacha Mytton and Doug Millar are employees of Genetic Signatures Limited. Sacha Mytton reports a relationship with Genetic Signatures Limited that includes: employment. Doug Millar reports a relationship with Genetic Signatures Limited that includes: employment. Co-authors Anuk Indraratna and Alison Ricafrente were previously employed by Genetic Signatures Limited. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
- Published
- 2024
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