دورية أكاديمية
Express Yourself: Quantitative Real-Time PCR Assays for Rapid Chromosomal Antimicrobial Resistance Detection in Pseudomonas aeruginosa.
| العنوان: | Express Yourself: Quantitative Real-Time PCR Assays for Rapid Chromosomal Antimicrobial Resistance Detection in Pseudomonas aeruginosa. |
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| المؤلفون: | Madden DE; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Olagoke O; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Baird T; Sunshine Coast Health Institute, Birtinya, Queensland, Australia.; Respiratory Department, Sunshine Coast University Hospital, Birtinya, Queensland, Australia., Neill J; Sunshine Coast Health Institute, Birtinya, Queensland, Australia.; Respiratory Department, Sunshine Coast University Hospital, Birtinya, Queensland, Australia., Ramsay KA; Child Health Research Centre, The University of Queenslandgrid.1003.2, South Brisbane, Queensland, Australia., Fraser TA; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Bell SC; Child Health Research Centre, The University of Queenslandgrid.1003.2, South Brisbane, Queensland, Australia.; Adult Cystic Fibrosis Centre, The Prince Charles Hospital, Chermside, Queensland, Australia.; Translational Research Institute, Woolloongabba, Queensland, Australia., Sarovich DS; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia., Price EP; Centre for Bioinnovation, University of the Sunshine Coastgrid.1034.6, Sippy Downs, Queensland, Australia.; Sunshine Coast Health Institute, Birtinya, Queensland, Australia. |
| المصدر: | Antimicrobial agents and chemotherapy [Antimicrob Agents Chemother] 2022 May 17; Vol. 66 (5), pp. e0020422. Date of Electronic Publication: 2022 Apr 25. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Society for Microbiology Country of Publication: United States NLM ID: 0315061 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1098-6596 (Electronic) Linking ISSN: 00664804 NLM ISO Abbreviation: Antimicrob Agents Chemother Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, American Society for Microbiology |
| مواضيع طبية MeSH: | Cystic Fibrosis*/complications , Pseudomonas Infections*/microbiology, Anti-Bacterial Agents/therapeutic use ; Bacterial Outer Membrane Proteins/genetics ; Bacterial Proteins/metabolism ; Carbapenems/therapeutic use ; Drug Resistance, Bacterial ; Humans ; Membrane Transport Proteins/genetics ; Microbial Sensitivity Tests ; Pseudomonas aeruginosa ; Real-Time Polymerase Chain Reaction |
| مستخلص: | The rise of antimicrobial-resistant (AMR) bacteria is a global health emergency. One critical facet of tackling this epidemic is more rapid AMR diagnosis in serious multidrug-resistant pathogens like Pseudomonas aeruginosa. Here, we designed and then validated two multiplex quantitative real-time PCR (qPCR) assays to simultaneously detect differential expression of the resistance-nodulation-division efflux pumps MexAB-OprM, MexCD-OprJ, MexEF-OprN, and MexXY-OprM, the AmpC β-lactamase, and the porin OprD, which are commonly associated with chromosomally encoded AMR. Next, qPCRs were tested on 15 sputa from 11 participants with P. aeruginosa respiratory infections to determine AMR profiles in vivo . We confirmed multiplex qPCR testing feasibility directly on sputa, representing a key advancement in in vivo AMR diagnosis. Notably, comparison of sputa with their derived isolates grown in Luria-Bertani broth (±2.5% NaCl) or a 5-antibiotic cocktail showed marked expression differences, illustrating the difficulty in replicating in vivo expression profiles in vitro . Cystic fibrosis sputa showed significantly reduced mexE and mexY expression compared with chronic obstructive pulmonary disease sputa, despite harboring fluoroquinolone- and aminoglycoside-resistant strains, indicating that these loci do not contribute to AMR in vivo. oprD was also significantly downregulated in cystic fibrosis sputa, even in the absence of contemporaneous carbapenem use, suggesting a common adaptive trait in chronic infections that may affect carbapenem efficacy. Sputum ampC expression was highest in participants receiving carbapenems (6.7 to 15×), some of whom were simultaneously receiving cephalosporins, the latter of which would be rendered ineffective by the upregulated ampC . Our qPCR assays provide valuable insights into the P. aeruginosa resistome, and their use on clinical specimens will permit timely treatment alterations that will improve patient outcomes and antimicrobial stewardship measures. |
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| فهرسة مساهمة: | Keywords: AMR; Pseudomonas aeruginosa; ampC; antibiotic; antibiotic resistance; assay development; diagnostics; efflux pumps; gene expression; genomics; in vivo expression technology; qPCR |
| المشرفين على المادة: | 0 (Anti-Bacterial Agents) 0 (Bacterial Outer Membrane Proteins) 0 (Bacterial Proteins) 0 (Carbapenems) 0 (Membrane Transport Proteins) |
| تواريخ الأحداث: | Date Created: 20220425 Date Completed: 20220519 Latest Revision: 20240828 |
| رمز التحديث: | 20240828 |
| مُعرف محوري في PubMed: | PMC9112894 |
| DOI: | 10.1128/aac.00204-22 |
| PMID: | 35467369 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1098-6596 |
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| DOI: | 10.1128/aac.00204-22 |