MP02-08 RAPID DETECTION OF BACTERIAL UTIS VIA REPORTER PHAGE-BASED BIOLUMINESCENCE ASSAY

You have accessJournal of UrologyCME1 May 2022MP02-08 RAPID DETECTION OF BACTERIAL UTIS VIA REPORTER PHAGE-BASED BIOLUMINESCENCE ASSAY Lorenz Leitner, Jiemin Du, Susanne Meile, Shawna McCallin, Vera Neumeier, Thomas M. Kessler, Martin J. Loessner, Samuel Kilcher, and Matthew Dunne Lorenz LeitnerLorenz Leitner More articles by this author , Jiemin DuJiemin Du More articles by this author , Susanne MeileSusanne Meile More articles by this author , Shawna McCallinShawna McCallin More articles by this author , Vera NeumeierVera Neumeier More articles by this author , Thomas M. KesslerThomas M. Kessler More articles by this author , Martin J. LoessnerMartin J. Loessner More articles by this author , Samuel KilcherSamuel Kilcher More articles by this author , and Matthew DunneMatthew Dunne More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002514.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Bacterial urinary tract infections (UTIs) are among the most common microbial diseases in all age groups and major contributors to the injudicious use of antibiotics. Rapid and accurate pathogen identification directly from urine samples can drastically improve the clinical management of UTIs, alleviating their socio-economic impacts and facilitating antibiotic stewardship. METHODS: Luciferase reporter phages are genetically engineered viruses capable of rapidly detecting their target bacterial hosts with exquisite specificity and sensitivity. As signal production requires phage genome injection and subsequent host-dependent expression of phage-encoded reporter proteins, this diagnostic method does not produce false-positive results in the presence of dead target cells. Here, using a set of six genetically distinct luciferase reporter phages, we report the development and evaluation of a novel phage-based bioluminescence assay for rapid detection of the three predominant UTI pathogens, Escherichia coli, Klebsiella spp. and Enterococcus spp. RESULTS: In vitro, with pure cultures, the assay positively identified 89% of 155 relevant clinical isolates, showing the highest inclusivity for Enterococcus spp. (50/52), followed by Klebsiella spp. (45/51) and E. coli (43/52). Cross-reactivity of the phage cocktails with other closely related genus including Raoultella and Staphylococcus was observed, but only at minimal rate (4.3%; 3/70). The assay performance was further assessed alongside routine clinical testing on 145 patient specimens collected from a local hospital in Switzerland over 5 months. Following short pre-enrichment steps and with simple substrate addition, the assay reliably detected the three target pathogens with 73% sensitivity and 96% specificity at a resolution of ≥103 CFU/ml in less than 6 h. CONCLUSIONS: This phage-based diagnostic platform offers opportunities for prompt bacterial UTI diagnosis in point-of-care settings and could be harnessed as companion diagnostics to assist rapid identification of potential responder patients during phage therapy. Source of Funding: Swiss National Science Foundation © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e13 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Lorenz Leitner More articles by this author Jiemin Du More articles by this author Susanne Meile More articles by this author Shawna McCallin More articles by this author Vera Neumeier More articles by this author Thomas M. Kessler More articles by this author Martin J. Loessner More articles by this author Samuel Kilcher More articles by this author Matthew Dunne More articles by this author Expand All Advertisement PDF DownloadLoading ...