دورية أكاديمية
Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye.
| العنوان: | Monitoring phage-induced lysis of gram-negatives in real time using a fluorescent DNA dye. |
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| المؤلفون: | Egido JE; Medical Microbiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Toner-Bartelds C; Medical Microbiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Costa AR; Department of Bionanoscience, Delft University of Technology, Delft, The Netherlands.; Kavli Institute of Nanoscience, Delft, The Netherlands.; Fagenbank, Delft, The Netherlands., Brouns SJJ; Department of Bionanoscience, Delft University of Technology, Delft, The Netherlands.; Kavli Institute of Nanoscience, Delft, The Netherlands.; Fagenbank, Delft, The Netherlands., Rooijakkers SHM; Medical Microbiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Bardoel BW; Medical Microbiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands., Haas PJ; Medical Microbiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands. p.j.a.haas@umcutrecht.nl. |
| المصدر: | Scientific reports [Sci Rep] 2023 Jan 16; Vol. 13 (1), pp. 856. Date of Electronic Publication: 2023 Jan 16. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Nature Publishing Group Country of Publication: England NLM ID: 101563288 Publication Model: Electronic Cited Medium: Internet ISSN: 2045-2322 (Electronic) Linking ISSN: 20452322 NLM ISO Abbreviation: Sci Rep Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: London : Nature Publishing Group, copyright 2011- |
| مواضيع طبية MeSH: | Fluorescent Dyes* , Bacteriophages*/genetics, Bacteria ; Anti-Bacterial Agents ; DNA |
| مستخلص: | Bacteriophages (phages) are viruses that specifically attack bacteria. Their use as therapeutics, which constitutes a promising alternative to antibiotics, heavily relies on selecting effective lytic phages against the pathogen of interest. Current selection techniques are laborious and do not allow for direct visualization of phage infection dynamics. Here, we present a method that circumvents these limitations. It can be scaled for high-throughput and permits monitoring of the phage infection in real time via a fluorescence signal readout. This is achieved through the use of a membrane-impermeant nucleic acid dye that stains the DNA of damaged or lysed bacteria and new phage progeny. We have tested the method on Pseudomonas aeruginosa and Klebsiella pneumoniae and show that an increase in fluorescence reflects phage-mediated killing. This is confirmed by other techniques including spot tests, colony plating, flow cytometry and metabolic activity measurements. Furthermore, we illustrate how our method may be used to compare the activity of different phages and to screen the susceptibility of clinical isolates to phage. Altogether, we present a fast, reliable way of selecting phages against Gram-negative bacteria, which may be valuable in optimizing the process of selecting phages for therapeutic use. (© 2023. The Author(s).) |
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| المشرفين على المادة: | 0 (Fluorescent Dyes) 0 (Anti-Bacterial Agents) 9007-49-2 (DNA) |
| تواريخ الأحداث: | Date Created: 20230116 Date Completed: 20230118 Latest Revision: 20240911 |
| رمز التحديث: | 20240911 |
| مُعرف محوري في PubMed: | PMC9842612 |
| DOI: | 10.1038/s41598-023-27734-w |
| PMID: | 36646746 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 2045-2322 |
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| DOI: | 10.1038/s41598-023-27734-w |