4.4 Article

Detection of somatic coliphages through a bioluminescence assay measuring phage mediated release of adenylate kinase and adenosine 5′-triphosphate

Journal

JOURNAL OF VIROLOGICAL METHODS
Volume 161, Issue 1, Pages 107-113

Publisher

ELSEVIER
DOI: 10.1016/j.jviromet.2009.05.021

Keywords

Rapid assay; Coliphages; Adenylate kinase; Adenosine 5 '-triphosphate; Bioluminescence

Funding

  1. Ministerio de Medio Ambiente [A10212007/1-04.2]
  2. Ministerio de Educacion y Ciencia [CTM 2005-02513, BES-2003-0563]
  3. Generalitat de Catalunya [2005SGR00592]
  4. Spanish Agency for International Cooperation (MAE-AECI)

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The feasibility of detecting somatic coliphages by phage infection of Escherichia coli WG5 and measurement of phage propagation by the lysis mediated release of the bacterial host adenylate kinase (AK) and adenosine T-triphosphate (ATP) detected by a bioluminescent signal was evaluated. After 2 h of incubation, all cultures infected with reference bacteriophage phi X174 showed a significant increase in the bioluminescent signal, even with number of phages as low as less of 10 plaque forming units (PFU). Naturally occurring somatic coliphages ensured a significant bioluminescent signal after 3 h of infection when >10 PFU were inoculated. These results indicate that an easy and reliable method to detect low numbers of coliphages in less than 3 h is feasible. (C) 2009 Elsevier B.V. All rights reserved.

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