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.
摘要：
化脓性链球菌(A组链球菌;GAS)是导致大量人类死亡率和发病率的革兰氏阳性细菌。GAS咽炎的常规诊断依赖于咽拭子培养，低吞吐量，慢,和相对侵入性的“黄金标准”。虽然分子方法越来越多地被利用，唾液作为GAS感染诊断液的潜力在很大程度上仍未被探索.这里,我们提出了一部小说，高通量，敏感,和强大的speBqPCR检测，使用创新的3base™技术可靠地检测唾液中的GAS（GeneticSignaturesLimited，悉尼,澳大利亚)。该测定已在基线上验证，急性,和从控制的人类感染疫苗接种链球菌（CHIVAS-M75）试验产生的恢复期唾液样本，其中健康的成人参与者接受emm75GAS的挑战。在这些定义明确的样本中，我们的高通量检测方法分别优于唾液中的咽喉培养和常规qPCR，确认3base™平台的实用性，证明了唾液作为诊断生物流体的可行性，并为开发用于检测GAS和其他口咽病原体的新型非侵入性方法铺平了道路。
