Web of Science: 30 cites, Scopus: 32 cites, Google Scholar: cites
Fast phage detection and quantification : An optical density-based approach
Rajnovic, Denis (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia)
Muñoz Berbel, Xavier (Centro Nacional de Microelectrónica. Instituto de Microelectrónica de Barcelona)
Mas Gordi, Jordi (Universitat Autònoma de Barcelona. Departament de Genètica i de Microbiologia)

Data: 2019
Resum: Since 1959 with the proposal of Double Agar Layer (DAL) method for phage detection and quantification, many sophisticated methods have emerged meanwhile. However, many of them are either too complex/expensive or insensitive to replace routine utilization of DAL method in clinical, environmental and industrial environments. For that purpose, we have explored an alternative method for the detection and quantification of bacteriophages that fulfills the criteria of being rapid, simple and inexpensive. In this paper we have developed a method based on the analysis of optical density kinetics in bacterial cultures exposed to phage-containing samples. Although the decrease in optical density caused by cell lysis was one of the first observable consequences of the effect of viral infection in bacterial cultures, the potential of the method for the assessment of phage abundance has never been fully exploited. In this work we carry out a detailed study of optical density kinetics in phage-infected bacterial cultures, as a function of both, phage abundance and initial concentration of the host organisms. In total, 90 different combinations of bacteria/phage concentrations have been used. The data obtained provide valuable information about sensitivity ranges, duration of the assay, percentages of inhibition and type of lysing behavior for each phage concentration. The method described can detect, as few as 10 phage particles per assay volume after a phage incubation period of 3. 5h. The duration of the assay can be shortened to 45min at the expense of losing sensitivity and increasing the limit of detection to 10 pfu/ ml. Despite using non-sophisticated technology, the method described has shown sensitivity and response time comparable to other high-end methods. The simplicity of the technology and of the analytical steps involved, make the system susceptible of miniaturization and automation for high-throughput applications which can be implemented in routine analysis in many environments.
Ajuts: Ministerio de Economía y Competitividad CTQ2014-54553-C3-2-R
Ministerio de Economía y Competitividad RTC-2016-5766-2
European Commission 720851
Drets: Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original. Creative Commons
Llengua: Anglès
Document: Article ; recerca ; Versió publicada
Matèria: Bacteriophage T4 ; Biological Assay ; Escherichia coli ; Kinetics
Publicat a: PloS one, Vol. 14, Issue 5 (May 2019) , p. e0216292, ISSN 1932-6203

DOI: 10.1371/journal.pone.0216292
PMID: 31071103

14 p, 1.5 MB

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