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flow-cytometry immunoassay for detection of residues in food

Identification

Key words competitive fluorescence immunoassay, flow cytometry, microsphere, drug residue, pesticide, food, fluorophore, bead
Latest version 2011/01/03
Completed by TTZ

How does it work?

Primary objective simple and fast method for analysis of low-molecular-weight compounds in food
Working principle The method describes a microsphere-based competitive fluorescence immunoassay. To that end, the antigen is bound to microspheres (beads) which are internally dyed with a fluorophore. These conjugates are incubated in a solution of the sample and antibodies. Those antibodies which did not bind to residues in the sample bind to the antigen bound to the microspheres. The sample with the residue-antibody-complex is washed out. After staining of the antibody bound to the microspheres with a fluorophore bound to a secondary antibody the percentage of bound antibodies can be detected via flow cytometric analysis. A dual laser enables identification of colour-coded beads as well as quantification of the fluorophore bound to the beads. The detected signal is conversely correlated with the residue concentration in the sample.

File:Flow-cytometry immunoassay.pdf

Images
Additional effects
  • low trace sample consumption
  • low costs due to small amount of immunoassay reagents
  • time-saving
  • higher sensitivity, greater dynamic range and reproducibility than conventional ELISA
  • capability for multiresidue analysis because each analyte can be encoded by one set of microspheres
  • automation is possible for parallel and high-throughput detection
Important process parameters
  • functionality of the beads for antigen immobilization
  • bead concentration
  • incubation time
  • primary and secondary antibody concentration
  • concentration of fluorophore-conjugated reporter antibody
Important product parameters

What can it be used for?

Products liquids
Operations residue analysis, quality assurance
Solutions for short comings reduction of food contamination, detection of contaminants (in an early stage of the product chain)

What can it NOT be used for?

Products non-liquids, only with previous extraction and clean-up
Operations
Other limitations extraction can be time consuming and costly depending on the food matrix
Risks or hazards no risks

Implementation

Maturity lab scale, tested with different food matrices
Modularity /Implementation technical equipment for flow cytometry
Consumer aspects Low-molecular-weight compounds like residues of animal drugs and pesticides can be harmful for human beings. Thus, a fast and easy method is necessary to enable widespread analysis of these compounds to protect consumers and enforce food law.
Legal aspects Groups of substances to be monitored as well as official sampling procedures are laid down in Directive 96/23/EC, Commission Decision 97/747/EC, Commission Decision 98/179/EC and Regulation (EC) No 396/2005.
Environmental aspects Low reagent consumption

Further Information

Institutes UPV, RIKILT, IRAS, ICT Prague
Companies Applied Cytometry Systems
References 1. Cháfer-Pericás, C., Maquieira, A., Puchades, R. (2010) Fast screening methods to detect antibiotic residues in food samples. Trends in Analytical Chemistry, 29, 9, 1038-1049

2. De Keizer, W., Bienenmann-Ploum, M., Bergwerff, A., Haasnoot, W. (2008) Flow cytometric immunoassay for sulfonamides in raw milk. Analytica Chimica Acta, 620, 1-2, 142-149

3. Hu, L., Zuo, P., Ye, B. (2010) Multicomponent mesofluidic system for the detection of veterinary drug residues based on competitive immunoassay. Analytical Biochemistry, 405, 1, 89-95

4. Meimaridou, A., Haasnoot, W., Noteboom, L., Mintzas, D., Pulkrabova, J., Hajslova, J., Nielen, M. (2010). Color encoded microbeads-based flow cytometric immunoassay for polycyclic aromatic hydrocarbons in food. Analytica Chimica Acta 672, 9–14.

5. Zou, M., Gao, H., Li, J., Xu, F., Wang, L., Jiang, J. (2008) Rapid determination of hazardous compounds in food based on a competitive fluorescence microsphere immunoassay. Analytical Biochemistry, 374, 2, 318-324

6. COUNCIL DIRECTIVE 96/23/EC of 29 April 1996 on measures to monitor certain substances and residues thereof in live animals and animal products. Official Journal L 125 , 23/05/1996 P. 0010 – 0032

7. 97/747/EC: Commission Decision of 27 October 1997 fixing the levels and frequencies of sampling provided for by Council Directive 96/23/EC for the monitoring of certain substances and residues thereof in certain animal products. Official Journal L 303 , 06/11/1997 P. 0012 – 0015

8. 98/179/EC: Commission Decision of 23 February 1998 laying down detailed rules on official sampling for the monitoring of certain substances and residues thereof in live animals and animal products. Official Journal L 065 , 05/03/1998 P. 0031 – 0034

9. Regulation (EC) No 396/2005 of the European Parliament and of the Council of 23 February 2005 on maximum residue levels of pesticides in or on food and feed of plant and animal origin and amending Council Directive 91/414/EEC. Official Journal L 070 , 16/03/2005 P. 0001 - 0016

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  • bead concentration
  • incubation time
  • primary and secondary antibody concentration
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  • functionality of the beads for antigen immobilization
  • bead concentration
  • incubation time
  • primary and secondary antibody concentration
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Analytical instruments 2.1.3 biological not applicable biotechnology ScienceDirect (key words: detection AND residues AND food AND immunoassay) (key words: cytometric microsphere immunoassay AND food) WikiSysop :Template:Review document :Template:Review status



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Created by Ttz on 3 January 2011, at 13:53