title: Bacterial detection using unlabeled phage amplification and mass spectrometry through structural and nonstructural phage markers.
abstract: According to the World Health Organization, food safety is an essential public health priority. In this context, we report a relevant proof of feasibility for the indirect specific detection of bacteria in food samples using unlabeled phage amplification coupled to ESI mass spectrometry analysis and illustrated with the model phage systems T4 and SPP1. High-resolving power mass spectrometry analysis (including bottom-up and top-down protein analysis) was used for the discovery of specific markers of phage infection. Structural components of the viral particle and nonstructural proteins encoded by the phage genome were identified. Then, targeted detection of these markers was performed on a triple quadrupole mass spectrometer operating in the selected reaction monitoring mode. E. coli at 1 × 10(5), 5 × 10(5), and 1 × 10(6) CFU/mL concentrations was successfully detected after only a 2 h infection time by monitoring phage T4 structural markers in Luria-Bertani broth, orange juice, and French bean stew ("cassoulet") matrices. Reproducible detection of nonstructural markers was also demonstrated, particularly when a high titer of input phages was required to achieve successful amplification. This strategy provides a highly time-effective and sensitive assay for bacterial detection.
authors: bioMérieux S.A. , 376, Chemin de l'Orme, 69280 Marcy-l'Etoile, France.
authors: Armelle
authors: A
authors: Martelet
authors: Guillaume
authors: G
authors: L'Hostis
authors: Paulo
authors: P
authors: Tavares
authors: Sandrine
authors: S
authors: Brasilès
authors: François
authors: F
authors: Fenaille
authors: Christine
authors: C
authors: Rozand
authors: Alain
authors: A
authors: Theretz
authors: Gaspard
authors: G
authors: Gervasi
authors: Jean-Claude
authors: JC
authors: Tabet
authors: Eric
authors: E
authors: Ezan
authors: Christophe
authors: C
authors: Junot
authors: Bruno H
authors: BH
authors: Muller
authors: François
authors: F
authors: Becher
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001412:Bacillus subtilis / Q000302:isolation & purification* / Q000821:virology; D001628:Beverages / Q000032:analysis* / Q000382:microbiology; D032084:Citrus sinensis; D003090:Coliphages / Q000235:genetics*; D004926:Escherichia coli / Q000302:isolation & purification* / Q000821:virology; D005504:Food Analysis; D006801:Humans; D008242:Lysogeny; D008461:Meat Products / Q000032:analysis* / Q000382:microbiology; D008969:Molecular Sequence Data; D019151:Peptide Library; D021241:Spectrometry, Mass, Electrospray Ionization; D013552:Swine; D014764:Viral Proteins / Q000235:genetics
chemical_list: D019151:Peptide Library; D014764:Viral Proteins