An antibody microarray, in multiwell plate format, for multiplex screening of foodborne pathogenic bacteria and biomolecules

Gehring, Andrew G.; Albin, David M.; Reed, Sue A.; Shu-I Tu; Brewster, Jeffrey D.
May 2008
Analytical & Bioanalytical Chemistry;May2008, Vol. 391 Issue 2, p497
Academic Journal
Intoxication and infection caused by foodborne pathogens are important problems worldwide, and screening tests for multiple pathogens are needed because foods may be contaminated with multiple pathogens and/or toxic metabolites. We developed a 96-well microplate, multiplex antibody microarray method to simultaneously capture and detect Escherichia coli O157:H7 and Salmonella enterica serovar Typhimurium ( S. typhimurium), as well as a biomolecule (chicken immunoglobulin G or IgG employed as a proteinaceous toxin analog) in a single sample. Microarrayed spots of capture antibodies against the targeted analytes were printed within individual wells of streptavidin-coated polystyrene 96-multiwell microtiter plates and a sandwich assay with fluorescein- or Cy3-labeled reporter antibodies was used for detection. (Printing was achieved with a conventional microarray printing robot that was operated with custom-developed microplate arraying software.) Detection of the IgG was realized from ca. 5 to 25 ng/mL, and detection of E. coli O157:H7 and S. typhimurium was realized from ca. 106 to 109 and ca. 107 to 109 cells/mL, respectively. Multiplex detection of the two bacteria and the IgG in buffer and in culture-enriched ground beef filtrate was established with a total assay (including detection) time of ca. 2.5 h. Detection of S. typhimurium was largely unaffected by high concentrations of the other bacteria and IgG as well as the ground beef filtrate, whereas a small decrease in response was observed for E. coli O157:H7. The multiwell plate, multiplex antibody microarray platform developed here demonstrates a powerful approach for high-throughput screening of large numbers of food samples for multiple pathogens and toxins.


Related Articles

  • Concentrations of Escherichia coli and Genetic Diversity and Antibiotic Resistance Profiling of Salmonella Isolated from Irrigation Water, Packing Shed Equipment, and Fresh Produce in Texas. Duffy, E. A.; Lucia, L. M.; Kells, J. M.; Castillo, A.; Pillai, S. D.; Acuff, G. R. // Journal of Food Protection;Jan2005, Vol. 68 Issue 1, p70 

    Fresh produce has been repeatedly implicated as a vehicle in the transmission of foodborne gastroenteritis. In an effort to assess the risk factors involved in the contamination of fresh produce with pathogenic bacteria, a total of 1,257 samples were collected from cantaloupe, oranges, and...

  • Foodborne illnesses.  // Hospitality (11724285);Aug2007, Vol. 43 Issue 8, p30 

    The article discusses issues related to foodborne illnesses in New Zealand. It occurs when food contaminated with disease-causing living germs or their poisons is eaten. It is caused by bacteria which are able to multiply very fast in moist and warm conditions where they have suitable food...

  • Microbiological quality assessment in a fish processing plant at Mandapam, Ramanathapuram District. Prabakaran, P.; Kannan, K. Sendeesh; Anand, M.; Pradeepa, V. // Archives of Applied Science Research;2011, Vol. 3 Issue 2, p135 

    Foodborne illness resulting from the consumption of contaminated food and the major causative agents are pathogenic bacteria, fungi, viruses and parasites. Foodborne illness also arises from improper handling, preparation and food storage. The study is planned to evaluate the microbiological...

  • Rapid, Sensitive, and Simultaneous Detection of Three Foodborne Pathogens Using Magnetic Nanobead-Based Immunoseparation and Quantum Dot-Based Multiplex Immunoassay. Hong Wang; Yanbin Li; Wang, Andrew; Slavik, Michael // Journal of Food Protection;Dec2011, Vol. 74 Issue 12, p2039 

    Losses caused by foodborne diseases are enormous in terms of human life, illness, medical costs, and food product recalls. Rapid detection of multiple bacterial pathogens in foods is extremely important to ensure food safety. The objective of this research was to develop a multiplex immunoassay...

  • Reduced Detectability of Listeria monocytogenes in the Presence of Listeria innocua. Zitz, Ulrike; Zunabovic, Marija; Domig, Konrad J.; Wilrich, Peter-Theodor; Kneifel, Wolfgang // Journal of Food Protection;Aug2011, Vol. 74 Issue 8, p1282 

    Recent foodborne crises have demonstrated the importance of monitoring food safety. In terms of microbiological criteria, food safety requires the reliable detection of pathogens such as Listeria monocytogenes along the food chain by appropriate analytical methods. However, indications exist...

  • Listeria spp. E Listeria monocytogenes NA PRODUÇÃO DE SALSICHAS TIPO HOT DOG. Cesar, Alessandra Paro Rodrigues; De Mesquita, Albenones José; Prado, Cristiano Sales; Nunes, Iolanda Aparecida; De Almeida Filho, Edivaldo Sampaio // Ciência Animal Brasileira;2011, Vol. 12 Issue 2, p339 

    Listeria monocytogenes is widely distributed in the environment and it has been isolated from food that were associated to outbreaks of high lethality in many countries. Thus, this bacterium represents an important pathogen to the public health. Ready-to-eat products, like cooked stuffed food,...

  • Reevaluation of Epidemiological Criteria for Identifying Outbreaks of Acute Gastroenteritis Due to Norovirus: United States, 1998-2000. Turcios, Reina M.; Widdowson, Marc-Alain; Sulka, Alana C.; Mead, Paul S.; Glass, Roger I. // Clinical Infectious Diseases;4/1/2006, Vol. 42 Issue 7, p964 

    Background. Noroviruses are believed to be the most common etiologic agent of foodborne outbreaks of gastroenteritis, yet diagnostic tests for these agents are not readily available in the United States. In the absence of assays to detect norovirus, several clinical and epidemiologic...

  • Detection and antibiotic susceptibility profiles of Listeria monocytogenes in wildlife and water samples in Kubah National Park, Sarawak, Malaysia. Lesley, M. B.; Velnetti, L.; Fazira, A. A.; Kasing, A.; Samuel, L.; Micky, V.; Awang, A. S. A. H. // International Food Research Journal;2016, Vol. 23 Issue 1, p360 

    This study was conducted to detect the presence of Listeria monocytogenes (L. monocytogenes) and screen for its antibiotic susceptibility characteristic from wildlife and water samples at Kubah National Park, Sarawak, Malaysia. Samples collected were incubated and streaked on selective medium...

  • Food That's Fit to Eat.  // American Fitness;Mar/Apr2006, Vol. 24 Issue 2, p31 

    The article presents tips on lowering the risk of illness from foodborne bacteria. It would be better to avoid eating soft cheeses made from unpasteurized milk, such as brie and Camembert, blue-veined cheese or Mexican-style cheeses such as queso blanco. It is also suggested to prevent eating...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics