title: Study of effectiveness of bioluminescent reporter phage assay on Y. pseudotuberculosis strains.
abstract: The method describes the phage-mediated transduction of a bioluminescent phenotype to cultivated Y. pseudotuberculosis cells which are subsequently measured using a microplate luminometer. Reporter phage assay is rapid detection technique and its efficiency is not affected by presence of contaminating bacteria, no sample preparation is needed and it has the ability to test multiple samples simultaneously in a 96-well microtiter plate format. Experiments were performed to develop the rapid detection technique for Y. pseudotuberculosis strains and study the ability of a reporter Yersinia phage to confer a bioluminescent signal to Y. pseudotuberculosis strains under different environmental conditions (media, temperature, bacterial number) for detection. Further, to determine if the Yersinia phage can detect Y. pseudotuberculosis in presence of other bacterial species. The results revealed that the developed reporter phage assay is not effective against wide range of Y. pseudotuberculosis. Y. pseudotuberculosis could be rapidly detected within 30 minutes at 28°C. The reporter phage assay could detect luminescence within 45 minutes when the bacterial cells were at the minimal concentration 105 cells/mL. The optimal detectable concentrations were 106-107 cells/mL at 28 and 37°C. The reporter phage assay could detect Y. pseudotuberculosis within 30 minutes in presence of other enteric bacteria without selective enrichment. It should be noted that the Yersinia reporter phage is specific to Yersinia pestis strains and it can be used to detect Y. pseudotuberculosis when samples exclude the existence of Y. pestis strains. In the presented study this aspect was foreseen.
authors.forename: M R
authors.initials: MR
authors.lastname: Mitiashvili
mesh_terms: D001435:Bacteriophages / Q000235:genetics*; D017930:Genes, Reporter; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics*; D015011:Yersinia pseudotuberculosis / Q000235:genetics / Q000302:isolation & purification* / Q000821:virology; D015012:Yersinia pseudotuberculosis Infections / Q000175:diagnosis* / Q000235:genetics
chemical_list: D008164:Luminescent Proteins