caa a22 4500 606157913 CHVBK 20210128100616.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s10096-014-2246-0 doi (NATIONALLICENCE)springer-10.1007/s10096-014-2246-0 Isolation and development of bioluminescent reporter phages for bacterial dysentery [Elektronische Daten] [D. Schofield, D. Wray, I. Molineux] Shigellosis is a significant cause of morbidity and mortality worldwide, most notably amongst children. Moreover, there is a global increase in the occurrence of multidrug-resistant isolates, including the epidemic and pandemic Shigella dysenteriae type 1 strain. We developed a bioluminescent reporter phage assay to facilitate detection and simultaneously determine antibiotic susceptibility. A Shigella flexneri phage (Shfl25875) was isolated from environmental wastewater and characterized by DNA sequencing. Shfl25875 is T4-like, harbors a 169,062-bp genome, and grows on most (28/29) S. flexneri strains and all 12 S. dysenteriae type 1 strains tested. The genes encoding bacterial luciferase were integrated into the Shfl25875 genome to create a "light-tagged” phage capable of transducing a bioluminescent phenotype to infected cells. Shfl25875::luxAB rapidly detects cultured isolates with high sensitivity. Specificity experiments indicate that the reporter does not respond to Shigella boydii, non-type 1 S. dysenteriae strains, and most non-Shigella Enterobacteriaceae. Shfl25875::luxAB generates ampicillin and ciprofloxacin susceptibility profiles that are similar to the standard Clinical and Laboratory Standards Institute (CLSI) growth microdilution method, but in a significantly shorter time. In addition, the reporter phage detects Shigella in mock-infected stool. This new reporter phage shows promise as a tool for the detection of cultured isolates or complex clinical samples. Springer-Verlag Berlin Heidelberg, 2002 Schofield D. Guild BioSciences, 1313B Ashley River Road, 29407, Charleston, SC, USA aut Wray D. Guild BioSciences, 1313B Ashley River Road, 29407, Charleston, SC, USA aut Molineux I. Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, Austin, TX, USA aut European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/2(2015-02-01), 395-403 0934-9723 34:2<395 2015 34 10096 https://doi.org/10.1007/s10096-014-2246-0 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s10096-014-2246-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Schofield D. Guild BioSciences, 1313B Ashley River Road, 29407, Charleston, SC, USA aut NATIONALLICENCE 700 1- Wray D. Guild BioSciences, 1313B Ashley River Road, 29407, Charleston, SC, USA aut NATIONALLICENCE 700 1- Molineux I. Department of Molecular Biosciences, Institute for Cellular and Molecular Biology, University of Texas at Austin, 78712, Austin, TX, USA aut NATIONALLICENCE 773 0- European Journal of Clinical Microbiology & Infectious Diseases Springer Berlin Heidelberg 34/2(2015-02-01), 395-403 0934-9723 34:2<395 2015 34 10096