Monoclonal antibodies that react with live Listeria spp

Torensma, Ruurd; Visser, Maarten R.; Aarsman, C. J. M.; Poppelier, Miriam J.J.G.; Fluit, Ad C.; Verhoef, J.

doi:10.1128/aem.59.8.2713-2716.1993

Cited by 26 publications

(11 citation statements)

References 23 publications

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“…The reactivity patterns of these antibodies indicate that they are different in terms of specificity from MAbs to surface antigens of listeriae described by other investigators, which are Listeria genus specific (7,11) or specific for L. monocytogenes and one or more other Listeria species (3,30,31) or react preferentially (14) or exclusively (4) with L. monocytogenes strains of all serotypes.…”

Section: Discussionmentioning

confidence: 66%

Monoclonal antibodies with a high degree of specificity for Listeria monocytogenes serotype 4b

Kathariou

Mizumoto

Allen

et al. 1994

Appl Environ Microbiol

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Strains ofListeria monocytogenes serotype 4b account for a large fraction of sporadic listeriosis cases, as well as all major food-borne epidemics attributed to this pathogen. We have identified a set of three monoclonal antibodies which showed a high degree of specificity for strains of L. monocytogenes serotype 4b. Two of these antibodies (c74.33 and c74.180, isotypes immunoglobulin M [IgM] and IgG3, respectively) recognized all serotype 4b strains, whereas antibody c74.22 (isotype IgGl) failed to recognize certain epidemic-associated strains. The corresponding antigens were located on the surface of the bacteria and were expressed following bacterial growth in different media and over a wide range of temperatures (4, 22, and 37°C). Heating L. monocytogenes cells at 80, 90, or 100°C abolished reactivity for c74.22 but not for c74.33 MAb. These MAbs were negative for all of the non-Listeria strains tested, including representatives of several gram-negative and gram-positive species. The surface antigen recognized by c74.22 appeared to be associated with the ability of the bacteria to enter (invade) mammalian cells in culture.

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Section: Discussionmentioning

confidence: 66%

Monoclonal antibodies with a high degree of specificity for Listeria monocytogenes serotype 4b

Kathariou

Mizumoto

Allen

et al. 1994

Appl Environ Microbiol

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“…The overall stress conditions affected the antibody reactions to whole cells ranging from 8% to 59% for all the antibodies. In most enzyme-immunoassay applications (indirect or sandwich assay), the detection limit is 10 6 CFU ml )1 (Torensma et al 1993;Oladepo et al 2000;Valdivieso-Garcia et al 2001), and it is speculated that these levels of reduction in ELISA reactions, in some situations, will give false-negative results with cells at the lower detection range. However, the impact would be even more severe for biosensor applications designed to detect low number of cells (Abdel-Hamid et al 1999;Iqbal et al 2000;Leonard et al 2003;Geng et al 2004;Taitt et al 2004;Zhang and Ji 2004) when reduction in antibody reactions will certainly give false-negative results.…”

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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“…PCR-SSCP is reported to be capable of detecting single-base mutations in a fragment of a few hundred base pairs and seems to be a promising method for the differentiation of species. Because of the particular interest and expertise in our laboratory, we first focused on the applicability of PCR-SSCP for the detection and identification of Clostridium and Listeria species (10,24,26). Then we used 111 bacterial isolates belonging to 15 genera and 40 species to further test this technique ( Table 1).…”

Section: Resultsmentioning

confidence: 99%

Rapid identification of bacteria by PCR-single-strand conformation polymorphism

1994

Self Cite

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A new molecular biological approach for the identification of bacteria is described. This approach employs PCR of bacterial cell lysates with conserved primers located in the 16S rRNA sequence flanking a variable region, and analysis of the amplified product was based on the principle of single-strand conformation polymorphism (SSCP). The PCR product was denatured and separated on a nondenaturing polyacrylamide gel. SSCP patterns were detected by silver staining the nucleic acids. The mobility of the single-stranded DNA is sequence dependent and could be used to identify the unknown bacteria. Feasibility of the technique was demonstrated for a broad panel of gram-negative and gram-positive bacteria. We tested over 100 strains of bacteria representing 15 genera and 40 species. With the use of only two primer sets, P11P-P13P and ER10-ER11, we were capable to discriminate the tested species at the genus and species levels. Species-specific patterns were obtained for, e.g., Clostridium spp., Listeria spp., Pseudomonas spp., and Enterobacter spp. PCR-SSCP is a sensitive technique; e.g., the sensitivity obtained for Escherichia coli cells was 30 CFU. This technique is a simple and rapid method for the detection and identification of a wide spectrum of bacteria by whole-cell-based PCR amplification with the use of conserved primers and identification by nondenaturing gel electrophoresis.

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Monoclonal antibodies that react with live Listeria spp

Cited by 26 publications

References 23 publications

Monoclonal antibodies with a high degree of specificity for Listeria monocytogenes serotype 4b

Monoclonal antibodies with a high degree of specificity for Listeria monocytogenes serotype 4b

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

Rapid identification of bacteria by PCR-single-strand conformation polymorphism

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