Tags

Type your tag names separated by a space and hit enter

A rapid multiplexed chemiluminescent immunoassay for the detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes pathogen bacteria.
J Agric Food Chem. 2007 Jun 27; 55(13):4933-9.JA

Abstract

A simple and rapid multiplexed sandwich chemiluminescent enzyme immunoassay has been developed for the simultaneous detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes. To achieve the multiplexed detection of the four pathogens, a new polystyrene 96 well microtiter plate format has been designed, in which each main well contains four subwells in the bottom. The monoclonal antibodies specific for each bacteria were separately immobilized in each subwell. When the samples were added to the main wells, the bacteria able to specifically bind to the corresponding monoclonal antibody were captured in one of the four subwells. Subsequently, a mixture of peroxidase-labeled polyclonal antibodies against the four bacteria was added and the peroxidase activity of the bound polyclonal labeled antibodies in each well was measured by an enhanced luminol-based chemiluminescent cocktail using a low-light charge-coupled imaging device. The assay was simple and fast, and the limit of quantification was in the order of 104-105 CFU/mL for all bacterial species. The accuracy of the method, evaluated by comparison of the results with a conventional culturing methodology, was satisfactory, with recovery values ranging from 90 to 120%. This method can be used as a screening test to evaluate the presence of these pathogen bacteria in different foodstuffs.

Authors+Show Affiliations

Magliulo M
Department of Pharmaceutical Sciences, University of Bologna, Bologna, Italy.
Simoni P
No affiliation info available
Guardigli M
No affiliation info available
Michelini E
No affiliation info available
Luciani M
No affiliation info available
Lelli R
No affiliation info available
Roda A
No affiliation info available

MeSH

Escherichia coli O157Food MicrobiologyImmunoenzyme TechniquesListeria monocytogenesLuminescent MeasurementsSalmonella typhimuriumYersinia enterocolitica

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

17530767

Citation

Magliulo, Maria, et al. "A Rapid Multiplexed Chemiluminescent Immunoassay for the Detection of Escherichia Coli O157:H7, Yersinia Enterocolitica, Salmonella Typhimurium, and Listeria Monocytogenes Pathogen Bacteria." Journal of Agricultural and Food Chemistry, vol. 55, no. 13, 2007, pp. 4933-9.
Magliulo M, Simoni P, Guardigli M, et al. A rapid multiplexed chemiluminescent immunoassay for the detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes pathogen bacteria. J Agric Food Chem. 2007;55(13):4933-9.
Magliulo, M., Simoni, P., Guardigli, M., Michelini, E., Luciani, M., Lelli, R., & Roda, A. (2007). A rapid multiplexed chemiluminescent immunoassay for the detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes pathogen bacteria. Journal of Agricultural and Food Chemistry, 55(13), 4933-9.
Magliulo M, et al. A Rapid Multiplexed Chemiluminescent Immunoassay for the Detection of Escherichia Coli O157:H7, Yersinia Enterocolitica, Salmonella Typhimurium, and Listeria Monocytogenes Pathogen Bacteria. J Agric Food Chem. 2007 Jun 27;55(13):4933-9. PubMed PMID: 17530767.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - A rapid multiplexed chemiluminescent immunoassay for the detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes pathogen bacteria. AU - Magliulo,Maria, AU - Simoni,Patrizia, AU - Guardigli,Massimo, AU - Michelini,Elisa, AU - Luciani,Mirella, AU - Lelli,Rossella, AU - Roda,Aldo, Y1 - 2007/05/26/ PY - 2007/5/29/pubmed PY - 2007/8/19/medline PY - 2007/5/29/entrez SP - 4933 EP - 9 JF - Journal of agricultural and food chemistry JO - J Agric Food Chem VL - 55 IS - 13 N2 - A simple and rapid multiplexed sandwich chemiluminescent enzyme immunoassay has been developed for the simultaneous detection of Escherichia coli O157:H7, Yersinia enterocolitica, Salmonella typhimurium, and Listeria monocytogenes. To achieve the multiplexed detection of the four pathogens, a new polystyrene 96 well microtiter plate format has been designed, in which each main well contains four subwells in the bottom. The monoclonal antibodies specific for each bacteria were separately immobilized in each subwell. When the samples were added to the main wells, the bacteria able to specifically bind to the corresponding monoclonal antibody were captured in one of the four subwells. Subsequently, a mixture of peroxidase-labeled polyclonal antibodies against the four bacteria was added and the peroxidase activity of the bound polyclonal labeled antibodies in each well was measured by an enhanced luminol-based chemiluminescent cocktail using a low-light charge-coupled imaging device. The assay was simple and fast, and the limit of quantification was in the order of 104-105 CFU/mL for all bacterial species. The accuracy of the method, evaluated by comparison of the results with a conventional culturing methodology, was satisfactory, with recovery values ranging from 90 to 120%. This method can be used as a screening test to evaluate the presence of these pathogen bacteria in different foodstuffs. SN - 0021-8561 UR - https://www.unboundmedicine.com/medline/citation/17530767/A_rapid_multiplexed_chemiluminescent_immunoassay_for_the_detection_of_Escherichia_coli_O157:H7_Yersinia_enterocolitica_Salmonella_typhimurium_and_Listeria_monocytogenes_pathogen_bacteria_ L2 - https://doi.org/10.1021/jf063600b DB - PRIME DP - Unbound Medicine ER -