Development of a monoclonal sandwich ELISA for the detection of animal and human Escherichia coli O157 strains

Kerr, P.; Chart, Henrik; Finlay, D.; Pollock, D.A.; Mackie, D.P.; Ball, H.J.

doi:10.1046/j.1365-2672.2001.01281.x

Cited by 37 publications

(28 citation statements)

References 41 publications

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“…The sandwich ELISA detection limit (10 5 CFU ml (1 ) is similar to most ELISAs already proposed for the detection of pathogenic bacteria in foods (Kerr et al, 2001;Westerman, He, Keen, Littledike, & Kwang, 1997). The ability of the sandwich ELISA was further investigated using the MAbs to detect Salmonella Typhimurium in experimentally contaminated food samples (beef and chicken meat) in the presence of their natural microbiota.…”

Section: Discussionsupporting

confidence: 60%

Monoclonal antibodies against serogroup B salmonellae: production, characterisation and use in a sandwich ELISA

Moreira

Conceição

et al. 2008

Food and Agricultural Immunology

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Ten monoclonal antibodies (MAbs) against serogroup B salmonellae were obtained after immunisation of BALB/c mice with outer membrane proteins (OMPs) from Salmonella enterica serovar Typhimurium. Affinity constants, measured by enzyme-linked immunosorbent assay (ELISA), ranged from 8 )0 5 to 6)0 7 l mol (1 . Additivity ELISA demonstrated that most MAbs recognise different epitopes. ELISA determined the antigen specificity of the MAbs. The MAbs were more reactive with live Salmonella Typhimurium than with heattreated cells. Two MAbs were used to develop a sandwich ELISA for rapid detection of Salmonella Typhimurium in experimentally contaminated meats. Its detection limit was 10 5 CFU ml (1 and it was able to detect 1Á10 CFU in postenrichment broth from 25 g of beef and chicken meat samples. The sandwich ELISA developed was shown to be specific and sensitive for the detection of Salmonella Typhimurium in meat, and produced results comparable to the culture methods in 57 h.

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Section: Discussionsupporting

confidence: 60%

Monoclonal antibodies against serogroup B salmonellae: production, characterisation and use in a sandwich ELISA

Moreira

Conceição

et al. 2008

Food and Agricultural Immunology

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“…In some studies using mAb as both capture and detection antibodies, the detection of the bacteria is often poor due to single targeting of the bacteria antigen. For instance, a sandwich ELISA using a mAb to the LPS of E. coli O157 has a detection limit of 10 5 CFU/ml [3]. Thus, we hypothesize the use of IgY may improve the bacteria detection because IgY is polyclonal, and therefore able to bind to multiple antigens on a single bacterium.…”

Section: Resultsmentioning

confidence: 99%

Detection of Escherichia coli O157:H7 using chicken immunoglobulin Y

2006

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“…Immunology-based assays have been developed for measuring specific antigens generally expressed on the outer membrane and secreted proteins from EHEC O157:H7. Kerr’s group (2001) developed a sandwich ELISA based on long-chain lipopolysaccharide to assay EHEC O157 from clinical samples, but cross reactivity with other bacteria occurred [16]. Here, we produced a polyclonal (pAb) and monoclonal antibody (mAb) against Intimin γ1 for the purpose of developing a sandwich ELISA to measure EHEC O157:H7 in clinical samples.…”

Section: Introductionmentioning

confidence: 99%

Development of a Sandwich ELISA for EHEC O157:H7 Intimin γ1

Zhang

et al. 2016

PLoS ONE

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Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a zoonotic pathogen of worldwide importance that causes foodborne infections in humans. Intimin gamma 1 (intimin γ1) is one of the most important outer membrane proteins required for EHEC’s intimate adhesion to epithelial cells. Here, we generated a polyclonal antibody (pAb) and a monoclonal antibody (mAb) against intimin γ1 to develop a double antibody sandwich ELISA (DAS-ELISA) with increased sensitivity and specificity for measuring EHEC O157:H7. To achieve this goal, a rabbit pAb was used as a capture antibody, and a mouse mAb was a detection antibody. No cross-reactivity was observed with the other genera of pathogenic bacteria tested with the DAS-ELISA, which included Salmonella enteritidis, Shigella flexneri type 2, Listeria monocytogenes, Streptococcus suis type 2, and other 18 serotype E. coli. Detection limits of the DAS-ELISA were 1 × 103 CFU/mL for EHEC O157:H7 cultures, 1 × 104 CFU/g before enrichment, and 1 × 102 CFU/g after enrichment of contaminated samples. Field samples (n = 498) were tested using a previously established duplex-PCR method and compared to our DAS-ELISA. The DAS-ELISA had a specificity of 94.4%, a sensitivity of 91.5% and accuracy of 94.0% compared with duplex-PCR. The DAS-ELISA developed here can be applied to EHEC O157:H7 quantification in food, animal, and environmental samples.

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Development of a monoclonal sandwich ELISA for the detection of animal and human Escherichia coli O157 strains

Cited by 37 publications

References 41 publications

Monoclonal antibodies against serogroup B salmonellae: production, characterisation and use in a sandwich ELISA

Monoclonal antibodies against serogroup B salmonellae: production, characterisation and use in a sandwich ELISA

Detection of Escherichia coli O157:H7 using chicken immunoglobulin Y

Development of a Sandwich ELISA for EHEC O157:H7 Intimin γ1

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