Detection of live and heat‐treated<i>Salmonella enteritidis</i>by a D<sub>1</sub>‐serospecific anti‐lipopolysaccharide O‐9 monoclonal antibody

Masi, Amy W.; Zawistowski, Jerzy

doi:10.1080/09540109509354895

Cited by 8 publications

(8 citation statements)

References 26 publications

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“…2C), which was anticipated, since the reporter antibody (mAb-2F11) was raised specifically against S. Enteritidis serovar (Masi & Zawistowski, 1995). These data are also in agreement with our previous study (Valadez et al, 2009).…”

Section: Resultssupporting

confidence: 92%

“…For fiber optic immunosensor-based detection, an S. entericaspecific rabbit polyclonal pAb-anti-Salmonella antibody (Valadez et al, 2009) was used as the capture antibody, whereas rabbit pAb-3238 developed in our laboratory (unpublished data) and mouse anti-S. Enteritidis mAb-2F11 (Jaradat, Bzikot, Zawistowski, & Bhunia, 2004;Masi & Zawistowski, 1995) were used as reporter antibodies. The specificity of antibody combinations to detect the target pathogens (10 8 CFU/mL) and their limit of detection (LOD) were optimized using enzyme linked immunosorbent assay (ELISA) (data not shown).…”

Section: Antibodies Labeling and Fiber Optics Sensormentioning

confidence: 99%

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Fiber optic and light scattering sensors: Complimentary approaches to rapid detection of Salmonella enterica in food samples

et al. 2016

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Section: Resultssupporting

confidence: 92%

Section: Antibodies Labeling and Fiber Optics Sensormentioning

confidence: 99%

Fiber optic and light scattering sensors: Complimentary approaches to rapid detection of Salmonella enterica in food samples

et al. 2016

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“…After three washings with TBST (TBS with 0.1% tween 20), the membranes were incubated overnight with either mouse antisera raised against OMP extracts and diluted 1:25 000 in TBST buffer containing 1% gelatin or MAb 1D6 diluted 1:5 in the same buffer. After several washings with TBST, the blots were developed using alkaline phosphate-conjugated anti-mouse immunoglobulins (Masi and Zawistowski, 1995).…”

Section: Immunoblottingmentioning

confidence: 99%

“…Cells were diluted with carbonate buffer (pH 9.6) to 10 8 cells ml" 1 and resulting suspension (100 ^1 per well) was used to coat 96-well plates overnight at 4°C. Then, plates were developed essentially as described earlier (Masi and Zawistowski, 1995).…”

Section: Elisa Protocolmentioning

confidence: 99%

Antigenically stable 35 kDa outer membrane protein ofSalmonella

Jaradat

Zawistowski

1998

Food and Agricultural Immunology

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Outer membrane protein (OMP) extracts from Salmonella species representing seven serogroups were examined by SDS-PAGE followed by immunoblotting. Two major proteins with apparent molecular weight of 24 and 35 kDa were identified. The latter protein was present only in Salmonella. Immunoblotting with a monoclonal antibody (MAb) 1D6 (IgA) demonstrated the 35 kDa OMP contains an antigen common for all tested Salmonella species with the exception of atypical species such as S. arizona. The type of growth media had no effect on the antigenicity of 35 kDa protein. However, the electrophoretic appearance of the 35 kDa protein was influenced by heat-treatment. Analysis of OMP extracts by SDS-PAGE and immunoblotting without heat-treatment revealed that MAb 1D6 bound several isoforms of this protein, a major form at 28 kDa and about eight minor forms in the range between 34 and 40 kDa. None of these forms contained carbohydrate moieties that may be responsible for the polymorphic appearance of the protein. These forms were converted to a single form by heat-treatment at 100°C indicating that the 35 kDa OMP is most likely a heat-modifiable protein. Furthermore, extended heattreatment (121°C, 15 min) did not affect the antigenicity of the 35 kDa OMP. Electron microscopy studies revealed that the 35 kDa OMP is exposed on a surface of the bacterial cells, although the frequency of epitopes recognized by the Mab 1D6 was low as can be inferred from the low number of gold particles attached to the cells.

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“…The tyvelose residue is a side sugar attached to the trisaccharide backbone through ␣1,3 linkage. This particular (20). One of the other important features of the immunoassay is based on the distribution of LPS in the gelled egg matrix formed upon heating.…”

mentioning

confidence: 99%

Application of Rapid Dot Blot Immunoassay for Detection of Salmonella enterica Serovar Enteritidis in Eggs, Poultry, and Other Foods

Yoshimasu

Zawistowski

2001

Appl Environ Microbiol

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Salmonella enterica serovar Enteritidis was detected in artificially inoculated eggs within 24 h through a rapid monoclonal antibody-based dot blot immunoassay. Detection in poultry and other products required 28 h. Samples were directly enriched in homogenized egg without the need for pre-or postenrichment steps. Serovar Enteritidis was detected in the presence of other bacteria when outcompeted 1:400.Conventional methods for detection of Salmonella in foods are labor-intensive, time-consuming, and expensive. It has also been found that some of the routinely used selective enrichment broths are inhibitory towards Salmonella enterica serovar Enteritidis (28). Rapid methods based on principles such as membrane technology (11), latex agglutination (27), immunoassays (6,18,30), and immunomagnetic separation (8,9,19) have been developed. Methods employing PCR in combination with preenrichment broths (24,31,32), immunomagnetic separation (25, 26), or centrifugation (21, 22) are currently being developed.Immunologically based methods specific for serovar Enteritidis suffer from the same drawbacks as the above methods: one or more enrichment broths, or in some cases, postenrichment broths, are required. Cross-reactivity has also been observed with most monoclonal antibodies produced against serovar Enteritidis (17,18,27). This report describes the development of a rapid dot blot immunoassay for the detection of serovar Enteritidis in eggs, poultry, and other products.Large grade A eggs were scrubbed with 70% ethanol and opened aseptically. Eggs were mixed for 30 s using a stomacher lab blender 400 (Seward Laboratory, London, England) and were inoculated with either serovar Enteritidis phage type 1, 4, 8, 13, or 13a. After incubation, 25-ml portions were placed into 50-ml polypropylene tubes, and 0.1 volume of 15% sodium cholate in phosphate-buffered saline (PBS) (pH 7.2) was added. After being mixed, tubes were placed in boiling water for 10 min and cooled for 30 min at 4°C. Through heating, the egg mixture was solidified, forming a solid egg gel. After cooling, the gel was removed from the tube, and a sterile core borer (10-mm diameter) was used to create small cylindrical gels. The cylindrical gels were then cut into disks of 2 mm in thickness.Nitrocellulose strips (8.5 by 2.5 cm) were prewetted with PBS prior to use. Egg disks were placed on the membrane for 5 min, removed, and washed for 2 min in PBS. Strips were blocked for 45 min in 5% skim milk powder in Tris-buffered saline (pH 7.5) and incubated with a murine monoclonal antibody solution (tissue culture supernatant) for 45 min, followed by 1 h of incubation in biotinylated goat anti-mouse immunoglobulin G. Strips were incubated with streptavidinalkaline phosphatase for 1 h and developed with a BCIP (5-bromo-4-chloro-3-indolylphosphate)-nitroblue tetrazolium chloride substrate solution. Membranes were washed twice with Tris-buffered saline containing 0.05% Tween 20 for 2 min between each step. All steps were performed at room temperature.Bacterial cultures were...

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Detection of live and heat‐treatedSalmonella enteritidisby a D₁‐serospecific anti‐lipopolysaccharide O‐9 monoclonal antibody

Cited by 8 publications

References 26 publications

Fiber optic and light scattering sensors: Complimentary approaches to rapid detection of Salmonella enterica in food samples

Fiber optic and light scattering sensors: Complimentary approaches to rapid detection of Salmonella enterica in food samples

Antigenically stable 35 kDa outer membrane protein ofSalmonella

Application of Rapid Dot Blot Immunoassay for Detection of Salmonella enterica Serovar Enteritidis in Eggs, Poultry, and Other Foods

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Detection of live and heat‐treatedSalmonella enteritidisby a D1‐serospecific anti‐lipopolysaccharide O‐9 monoclonal antibody

Cited by 8 publications

References 26 publications

Fiber optic and light scattering sensors: Complimentary approaches to rapid detection of Salmonella enterica in food samples

Fiber optic and light scattering sensors: Complimentary approaches to rapid detection of Salmonella enterica in food samples

Antigenically stable 35 kDa outer membrane protein ofSalmonella

Application of Rapid Dot Blot Immunoassay for Detection of Salmonella enterica Serovar Enteritidis in Eggs, Poultry, and Other Foods

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Detection of live and heat‐treatedSalmonella enteritidisby a D₁‐serospecific anti‐lipopolysaccharide O‐9 monoclonal antibody