Unstable Expression and Thermal Instability of a Species-Specific Cell Surface Epitope Associated with a 66-Kilodalton Antigen Recognized by Monoclonal Antibody EM-7G1 within Serotypes of
            <i>Listeria monocytogenes</i>
            Grown in Nonselective and Selective Broths

Nannapaneni, Ramakrishna; Story, Robert; Bhunia, Arun K.; Johnson, Michael G.

doi:10.1128/aem.64.8.3070-3074.1998

Cited by 22 publications

(13 citation statements)

References 27 publications

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“…Selective enrichment broths have previously shown to severely affect the expression of anti‐ Listeria monoclonal and polyclonal antibodies (Nannapaneni et al. 1998a,b; Geng et al.…”

Section: Discussionmentioning

confidence: 99%

Differential expression of InlB and ActA in Listeria monocytogenes in selective and nonselective enrichment broths

Lathrop¹,

Banada²,

Bhunia

2008

J Appl Microbiol

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Aim: To investigate the effect of selective and nonselective media on the expression of ActA and InlB proteins in Listeria monocytogenes. Methods and Results: Polyclonal antibodies to InlB and ActA were used in western blotting to determine the effect of selective (BLEB, UVM, and FB) or nonselective (BHI and LB) enrichment broths or hotdog exudates. Of the 13 L. monocytogenes serotypes tested, 11 and 12 serotypes showed a strong InlB expression in brain heart infusion (BHI) and Luria‐Bertani (LB), respectively, while only seven and one serotypes showed a strong ActA expression in these two respective broths, and others showed a weaker or no expression. On the contrary, in selective broths, expression of InlB was either very weak or undetectable. However, ActA expression was stronger in 12 serotypes when grown in buffered Listeria enrichment broth (BLEB), 11 in University of Vermont medium (UVM), and 10 in Fraser broth (FB). When tested in hotdog exudates, InlB and ActA were detected in serotypes grown at 37°C but not at 4°C. Transmission electron microscopy, enzyme‐linked immunosorbent assay, and mRNA analysis further supported these observations. Conclusion: Overall, selective enrichment broths promote ActA while nonselective broths promote InlB expression. Significance and Impact of the study: As commonly recommended enrichment broths show differential InlB and ActA expression, proper media must be selected to avoid false results during antibody‐based detection of L. monocytogenes.

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“…Selective enrichment broths have previously shown to severely affect the expression of anti‐ Listeria monoclonal and polyclonal antibodies (Nannapaneni et al. 1998a,b; Geng et al.…”

Section: Discussionmentioning

confidence: 99%

Differential expression of InlB and ActA in Listeria monocytogenes in selective and nonselective enrichment broths

Lathrop¹,

Banada²,

Bhunia

2008

J Appl Microbiol

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“…The data are the average of three independent tests and bars marked with different letters (a, b) are significantly thereby the detection of pathogenic bacteria (Ozkanca and Flint 2002). In particular, antibody-based detection, in which the antibodies recognize cellular surface components, could be seriously affected by environmental stresses (Geng et al 2003) or selective agents in the enrichment media (Nannapaneni et al 1998;Geng et al 2004). Actually, there are many studies on the expression of genes or proteins under various stress conditions, such as rpoS regulon of E. coli (Loewen et al 1998), acid shock proteins of Salmonella (Foster 1991), and r B and its associated regulatory proteins of L. monocytogenes (Wiedmann et al 1998).…”

Section: Discussionmentioning

confidence: 99%

Effect of environmental stresses on antibody-based detection of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis and Listeria monocytogenes

Hahm

Bhunia

2006

J Appl Microbiol

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Aims: To study the reaction patterns of selected antibodies to Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis and Listeria monocytogenes cells exposed to various environmental stresses. Methods and Results: Escherichia coli O157:H7, Salmonella Enteritidis and L. monocytogenes cells subjected to different environmental stress of temperatures (4 and 45°C), NaCl (5·5%), oxidative stress (15 mmol−1 H2O2), acidic pH (5·5) and ethanol (5%) for 3 h (short‐term stress) or for 5 days (long‐term stress) were analysed by ELISA and Western blotting. The ELISA results indicated that most stresses caused 12–16% reductions in reaction for anti‐E. coli O157:H7 and 20–48% reductions for anti‐Salmonella polyclonal antibodies during short‐term stress, whereas the most stresses exhibited enhanced reaction (44–100% increase) with the anti‐L. monocytogenes polyclonal antibody. During long‐term stress exposure to combined stress conditions of pH 5·5, 3·5% NaCl at 12°C or at 4°C, antibody reactions to the three pathogens were highly variable with the combined stress at 4°C showing the most reductions (8–40%). Likewise, there were about 18–59% reductions in antibody reactions with pathogens when cultured in hotdog samples with the combined stress conditions. Western blot analyses of crude cell surface antigens from both short‐ and long‐term stressed cells revealed that the changes in antibody reactions observed in ELISA were either because of repression, expression or possible denaturation of antigens on the surface of cells. Conclusions: Overall, the antibody reactions were significantly reduced in pathogens exposed to both short‐ and long‐term environmental stresses in culture medium or in meat sample because of expression, repression or denaturation of specific antigens in cells. Significance and Impact of the Study: In order to ensure the reliable detection of foodborne pathogens using antibody‐based methods, the influence of stress on antibody reactions should be thoroughly examined and understood first as the physiological activities in cells are often altered in response to a stress.

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“…These data indicate that FB may not be suitable for aptamer-based fibre-optic sensor application. In a previous report, we have shown that antimicrobial agents in FB tend to suppress antigen expression (Nannapaneni et al 1998;Lathrop et al 2008) thus affecting L. monocytogenes detection by antigen-specific antibodies or other ligands (Nannapaneni et al 1998;Geng et al 2004;Kim and Bhunia 2008). Therefore, appropriate enrichment broth must be selected for each sensor application to avoid false-negative result.…”

Section: Discussionmentioning

confidence: 99%

Antibody-aptamer functionalized fibre-optic biosensor for specific detection of Listeria monocytogenes from food

Ohk

Koo

Sen

et al. 2010

Journal of Applied Microbiology

153

107

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Aim: To develop antibody–aptamer functionalized fibre‐optic biosensor for specific detection of Listeria monocytogenes from food products. Methods and Results: Aptamer, a single‐stranded oligonucleotide ligand that displays affinity for the target molecule, was used in the assay to provide sensor specificity. Aptamer‐A8, specific for internalin A, an invasin protein of L. monocytogenes, was used in the fibre‐optic sensor together with antibody in a sandwich format for detection of L. monocytogenes from food. Biotinylated polyclonal anti‐Listeria antibody, P66, was immobilized on streptavidin‐coated optical waveguide surface for capturing bacteria, and Alexa Fluor 647‐conjugated A8 was used as a reporter. The biosensor was able to selectively detect pathogenic Listeria in pure culture and in mixture with other bacteria at a concentration of approx. 103 CFU ml−1. This sensor also successfully detected L. monocytogenes cells from artificially contaminated (initial inoculation of 102 CFU 25 g−1) ready‐to‐eat meat products such as sliced beef, chicken and turkey after 18 h of enrichment. Conclusion: Based on the data presented in this study, the antibody–aptamer functionalized fibre‐optic biosensor could be used as a detection tool for sensitive and specific detection of L. monocytogenes from foods. Significance and Impact of the Study: The study demonstrates feasibility and novel application of aptamer on fibre‐optic biosensor platform for the sensitive detection of L. monocytogenes from food products.

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Unstable Expression and Thermal Instability of a Species-Specific Cell Surface Epitope Associated with a 66-Kilodalton Antigen Recognized by Monoclonal Antibody EM-7G1 within Serotypes of Listeria monocytogenes Grown in Nonselective and Selective Broths

Cited by 22 publications

References 27 publications

Differential expression of InlB and ActA in Listeria monocytogenes in selective and nonselective enrichment broths

Differential expression of InlB and ActA in Listeria monocytogenes in selective and nonselective enrichment broths

Effect of environmental stresses on antibody-based detection of Escherichia coli O157:H7, Salmonella enterica serotype Enteritidis and Listeria monocytogenes

Antibody-aptamer functionalized fibre-optic biosensor for specific detection of Listeria monocytogenes from food

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