Dataset Information

Flow cytometry-based rapid detection of Staphylococcus aureus and Pseudomonas aeruginosa using fluorescent antibodies.

ABSTRACT: Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) are major pathogens frequently detected in food and beverage poisoning, and persistent infections. Therefore, the development of a rapid method that can detect these pathogens before serious multiplication is required. In this study, we established a flow cytometry (FCM)-based detection method that allows rapid acquisition of cell populations in fluid samples by using a fluorescent antibody against S. aureus or P. aeruginosa. Using this method, we detected these pathogens with a 10³ to 10⁵ CFU order of limit of detection value within 1 hour. The FCM-based method for the detection of S. aureus and P. aeruginosa offers the possibility of high-throughput analysis of pathogens in food, environmental, and clinical sources.

SUBMITTER: Kim JK

PROVIDER: S-EPMC9717348 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Publications

Flow cytometry-based rapid detection of Staphylococcus aureus and Pseudomonas aeruginosa using fluorescent antibodies.

Kim Joo-Kyung JK Yun Hanool H Yeom Chang-Hun CH Kim Eun-Jung EJ Kim Wooseong W Lee Chang-Soo CS Kim Byung-Gee BG Jeong Hee-Jin HJ

RSC advances 20221101 53

Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) are major pathogens frequently detected in food and beverage poisoning, and persistent infections. Therefore, the development of a rapid method that can detect these pathogens before serious multiplication is required. In this study, we established a flow cytometry (FCM)-based detection method that allows rapid acquisition of cell populations in fluid samples by using a fluorescent antibody a ...[more]

PMID: 36545616

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Project description:Cystic fibrosis (CF) patients chronically infected with both Pseudomonas aeruginosa and Staphylococcus aureus have worse health outcomes than patients who are monoinfected with either P. aeruginosa or S. aureus We showed previously that mucoid strains of P. aeruginosa can coexist with S. aureusin vitro due to the transcriptional downregulation of several toxic exoproducts typically produced by P. aeruginosa, including siderophores, rhamnolipids, and HQNO (2-heptyl-4-hydroxyquinoline N-oxide). Here, we demonstrate that exogenous alginate protects S. aureus from P. aeruginosa in both planktonic and biofilm coculture models under a variety of nutritional conditions. S. aureus protection in the presence of exogenous alginate is due to the transcriptional downregulation of pvdA, a gene required for the production of the iron-scavenging siderophore pyoverdine as well as the downregulation of the PQS (Pseudomonas quinolone signal) (2-heptyl-3,4-dihydroxyquinoline) quorum sensing system. The impact of exogenous alginate is independent of endogenous alginate production. We further demonstrate that coculture of mucoid P. aeruginosa with nonmucoid P. aeruginosa strains can mitigate the killing of S. aureus by the nonmucoid strain of P. aeruginosa, indicating that the mechanism that we describe here may function in vivo in the context of mixed infections. Finally, we investigated a panel of mucoid clinical isolates that retain the ability to kill S. aureus at late time points and show that each strain has a unique expression profile, indicating that mucoid isolates can overcome the S. aureus-protective effects of mucoidy in a strain-specific manner.IMPORTANCE CF patients are chronically infected by polymicrobial communities. The two dominant bacterial pathogens that infect the lungs of CF patients are P. aeruginosa and S. aureus, with ∼30% of patients coinfected by both species. Such coinfected individuals have worse outcomes than monoinfected patients, and both species persist within the same physical space. A variety of host and environmental factors have been demonstrated to promote P. aeruginosa-S. aureus coexistence, despite evidence that P. aeruginosa kills S. aureus when these organisms are cocultured in vitro Thus, a better understanding of P. aeruginosa-S. aureus interactions, particularly mechanisms by which these microorganisms are able to coexist in proximal physical space, will lead to better-informed treatments for chronic polymicrobial infections.

Dataset Information

Flow cytometry-based rapid detection of Staphylococcus aureus and Pseudomonas aeruginosa using fluorescent antibodies.

Publications

Flow cytometry-based rapid detection of <i>Staphylococcus aureus</i> and <i>Pseudomonas aeruginosa</i> using fluorescent antibodies.

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