First Application of a Microsphere-Based Immunoassay to the Detection of Genetically Modified Organisms (GMOs):  Quantification of Cry1Ab Protein in Genetically Modified Maize

Fantozzi, Anna; Ermolli, Monica; Marini, Massimiliano; Scotti, Domenico; Balla, Branko; Querci, Maddalena; Langrell, Stephen; Eede, Guy Van den

doi:10.1021/jf061506p

Cited by 41 publications

(23 citation statements)

References 10 publications

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“…A number of researchers have performed microsphere-based assays to detect potential contaminants of foods: Dunbar et al [2] detected several bacterial pathogens in phosphate-buffered saline (PBS), Ikeda et al [3] detected pathogenic bacteria using extracted RNA, Fantozzi et al [4] detected a genetically modified maize protein, while Haasnoot and du Pre [5] demonstrated a triplex immunoassay for vegetable proteins in a milk powder matrix. Although these studies demonstrated detection of food-borne pathogens, with the exception of Haasnoot and du Pre, application of these assays to a wide range of real-world food matrices was not demonstrated.…”

Section: Introductionmentioning

confidence: 99%

Multiplexed magnetic microsphere immunoassays for detection of pathogens in foods

Kim

Taitt

Ligler

et al. 2010

Sens. & Instrumen. Food Qual.

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Foodstuffs have traditionally been challenging matrices for conducting immunoassays. Proteins, carbohydrates, and other macromolecules present in food matrices may interfere with both immunoassays and PCR-based tests, and removal of particulate matter may also prove challenging prior to analyses. This has been found true when testing for bacterial contamination of foods using the standard polystyrene microspheres utilized with Luminex flow cytometers. Luminex MagPlex microspheres are encoded with the same dyes as standard xMAP microspheres, but have superparamagnetic properties to aid in preparation of samples in complex matrices. In this work, we present results demonstrating use of MagPlex for sample preparation and identification of bacteria and a toxin spiked into a variety of food samples. Fluorescence-coded MagPlex microsphere sets coated with antibodies for Salmonella, Campylobacter, Escherichia coli, Listeria, and staphylococcal enterotoxin B (SEB) were used to capture these bacteria and toxin from spiked foodstuffs and then evaluated by the Luminex system in a multiplex format; spiked foods included apple juice, green pepper, tomato, ground beef, alfalfa sprouts, milk, lettuce, spinach, and chicken washes. Although MagPlex microspheres facilitated recovery of the microspheres and targets from the complex matrices, assay sensitivity was sometimes inhibited by up to one to three orders of magnitude; for example the detection limits E. coli spiked into apple juice or milk increased 100-fold, from 1000 to 100,000 cfu/mL. Thus, while the magnetic and fluorescent properties of the Luminex MagPlex microspheres allow for rapid, multiplexed testing for bacterial contamination in typically problematic food matrices, our data demonstrate that achieving desired limits of detection is still a challenge.

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Section: Introductionmentioning

confidence: 99%

Multiplexed magnetic microsphere immunoassays for detection of pathogens in foods

Kim

Taitt

Ligler

et al. 2010

Sens. & Instrumen. Food Qual.

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“…Detection of the novel proteins produced by GM crops relies almost exclusively on the application of immunoassay technology (Fantozzi et al, 2007). Commercial immunoassays are available for most of the GM crops on the market today and have been used in a variety of large-scale applications, determining GM content (%GM) ensuring compliance with non-GM labeling requirements, and confirming the presence of high-value commodities.…”

Section: Methods Based On Protein: Immunoassay Technologymentioning

confidence: 99%

Testing Methods for Agriculture and Food Safety

Jiménez-López¹,

Hernandez‐Soriano²

2012

Biochemical Testing

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“…The microsphere-based technology was also applied for the detection of Cry1Ab protein in genetically modified maize with an LOD of 0.018% (weight (GMO)/ weight), and was described as the first application of a quantitative high-throughput immunoassay in GMO analysis with multiplex options by Fantozzi et al [37].…”

Section: Food Allergens and Proteinaceous Adulterantsmentioning

confidence: 99%

Multiplex bioanalytical methods for food and environmental monitoring

Raz¹,

Haasnoot²

2011

TrAC Trends in Analytical Chemistry

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Recent advances in miniaturization of analytical systems and newly emerging technologies offer platforms with greater automation and multiplexing capabilities than traditional biological binding assays. Multiplexed bioanalytical techniques provide control agencies and food industries with new possibilities for improved, more efficient monitoring of food and environmental contaminants. This review deals with recent developments in planar-array and suspension-array technologies, and their applications in detecting pathogens, food allergens and adulterants, toxins, antibiotics and environmental contaminants. ª

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First Application of a Microsphere-Based Immunoassay to the Detection of Genetically Modified Organisms (GMOs): Quantification of Cry1Ab Protein in Genetically Modified Maize

Cited by 41 publications

References 10 publications

Multiplexed magnetic microsphere immunoassays for detection of pathogens in foods

Multiplexed magnetic microsphere immunoassays for detection of pathogens in foods

Testing Methods for Agriculture and Food Safety

Multiplex bioanalytical methods for food and environmental monitoring

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