title: Rapid Detection of Listeria by Bacteriophage Amplification and SERS-Lateral Flow Immunochromatography.
abstract: A rapid Listeria detection method was developed utilizing A511 bacteriophage amplification combined with surface-enhanced Raman spectroscopy (SERS) and lateral flow immunochromatography (LFI). Anti-A511 antibodies were covalently linked to SERS nanoparticles and printed onto nitrocellulose membranes. Antibody-conjugated SERS nanoparticles were used as quantifiable reporters. In the presence of A511, phage-SERS nanoparticle complexes were arrested and concentrated as a visible test line, which was interrogated quantitatively by Raman spectroscopy. An increase in SERS intensity correlated to an increase in captured phage-reporter complexes. SERS limit of detection was 6 × 10(6) pfu·mL(-1), offering detection below that obtainable by the naked eye (LOD 6 × 10(7) pfu·mL(-1)). Phage amplification experiments were carried out at a multiplicity of infection (MOI) of 0.1 with 4 different starting phage concentrations monitored over time using SERS-LFI and validated by spot titer assay. Detection of L. monocytogenes concentrations of 1 × 10(7) colony forming units (cfu)·mL(-1), 5 × 10(6) cfu·mL(-1), 5 × 10(5) cfu·mL(-1) and 5 × 10(4) cfu·mL(-1) was achieved in 2, 2, 6, and 8 h, respectively. Similar experiments were conducted at a constant starting phage concentration (5 × 10(5) pfu·mL(-1)) with MOIs of 1, 2.5, and 5 and were detected in 2, 4, and 5 h, respectively.
authors.affiliation: Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA. nstambac@mines.edu.
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Stambach
authors.affiliation: Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA. scarr@mines.edu.
authors.forename: Stephanie A
authors.initials: SA
authors.lastname: Carr
authors.affiliation: Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA. crcox@mines.edu.
authors.forename: Christopher R
authors.initials: CR
authors.lastname: Cox
authors.affiliation: Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA. kvoorhee@mines.edu.
authors.forename: Kent J
authors.initials: KJ
authors.lastname: Voorhees
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D001435:Bacteriophages / Q000254:growth & development* / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D003955:Diagnostic Tests, Routine / Q000379:methods*; D008087:Listeria / Q000821:virology*; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013997:Time Factors
keywords: A511; Listeria monocytogenes; lateral flow immunochromatography; phage amplification; surface-enhanced Raman spectroscopy