Rapid Detection of Listeria by Bacteriophage Amplification and SERS-Lateral Flow Immunochromatography
Department of Chemistry and Geochemistry, Colorado School of Mines, Golden, CO 80401, USA
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Author to whom correspondence should be addressed.
Academic Editors: Abram Aertsen and Rob Lavigne
Viruses 2015, 7(12), 6631-6641; https://doi.org/10.3390/v7122962
Received: 28 August 2015
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Revised: 2 November 2015
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Accepted: 10 December 2015
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Published: 14 December 2015
(This article belongs to the Special Issue 21st Evergreen International Phage Biology Meeting - Emerging Research)
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 × 106 pfu·mL−1, offering detection below that obtainable by the naked eye (LOD 6 × 107 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 × 107 colony forming units (cfu)·mL−1, 5 × 106 cfu·mL−1, 5 × 105 cfu·mL−1 and 5 × 104 cfu·mL−1 was achieved in 2, 2, 6, and 8 h, respectively. Similar experiments were conducted at a constant starting phage concentration (5 × 105 pfu·mL−1) with MOIs of 1, 2.5, and 5 and were detected in 2, 4, and 5 h, respectively.
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Keywords:
lateral flow immunochromatography; Listeria monocytogenes; surface-enhanced Raman spectroscopy; A511; phage amplification
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MDPI and ACS Style
Stambach, N.R.; Carr, S.A.; Cox, C.R.; Voorhees, K.J. Rapid Detection of Listeria by Bacteriophage Amplification and SERS-Lateral Flow Immunochromatography. Viruses 2015, 7, 6631-6641. https://doi.org/10.3390/v7122962
AMA Style
Stambach NR, Carr SA, Cox CR, Voorhees KJ. Rapid Detection of Listeria by Bacteriophage Amplification and SERS-Lateral Flow Immunochromatography. Viruses. 2015; 7(12):6631-6641. https://doi.org/10.3390/v7122962
Chicago/Turabian StyleStambach, Nicholas R., Stephanie A. Carr, Christopher R. Cox, and Kent J. Voorhees. 2015. "Rapid Detection of Listeria by Bacteriophage Amplification and SERS-Lateral Flow Immunochromatography" Viruses 7, no. 12: 6631-6641. https://doi.org/10.3390/v7122962
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