Multiplexed magnetic microsphere immunoassays for detection of pathogens in foods
Tóm tắt
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.
Tài liệu tham khảo
D.A. Vignali, Multiplexed particle-based flow cytometric assays. J. Immunol. Methods 243(1–2), 243–255 (2000)
S.A. Dunbar, C.A. Vander Zee, K.G. Oliver, K.L. Karem, J.W. Jacobson, Quantitative, multiplexed detection of bacterial pathogens: DNA and protein applications of the Luminex LabMAP system. J. Microbiol. Methods 53(2), 245–252 (2003)
M. Ikeda, N. Yamaguchi, K. Tani, M. Nasu, Rapid and simple detection of food poisoning bacteria by bead assay with a microfluidic chip-based system. J. Microbiol. Methods 67(2), 241–247 (2006)
A. Fantozzi, M. Ermolli, M. Marini, D. Scotti, B. Balla, M. Querci, S.R. Langrell, G. Van den Eede, First application of a microsphere-based immunoassay to the detection of genetically modified organisms (GMOs): quantification of Cry1Ab protein in genetically modified maize. J. Agric. Food Chem. 55(4), 1071–1076 (2007)
W. Haasnoot, J.G. du Pre, Luminex-based triplex immunoassay for the simultaneous detection of soy, pea, and soluble wheat proteins in milk powder. J. Agric. Food Chem. 55(10), 3771–3777 (2007)
K. Malkova, P. Rauch, G.M. Wyatt, M.R.A. Morgan, Combined immunomagnetic separation and detection of Salmonella enteritidis in food samples. Food Agric. Immunol. 10(3), 271–280 (1998)
J.H. Bergervoet, J. Peters, J.R. van Beckhoven, G.W. van den Bovenkamp, J.W. Jacobson, J.M. van der Wolf, Multiplex microsphere immuno-detection of potato virus Y, X and PLRV. J. Virol. Methods 149(1), 63–68 (2008)
C.R. Taitt, Y.S. Shubin, R. Angel, F.S. Ligler, Detection of Salmonella enterica serovar typhimurium by using a rapid, array-based immunosensor. Appl. Environ. Microbiol. 70(1), 152–158 (2004)
J.S. Kim, G.P. Anderson, J.S. Erickson, J.P. Golden, M. Nasir, F.S. Ligler, Multiplexed detection of bacteria and toxins using a microflow cytometer. Anal. Chem. 81(13), 5426–5432 (2009)
H.A. Waxman, R. DeLauro, FDA, Fresh Spinach Safety, Report to US House of Representatives Committee on Oversight and Government Reform 2008, 111th Congress, pp. 1–10
A.A. Ogunjimi, P.V. Choudary, Adsorption of endogenous polyphenols relieves the inhibition by fruit juices and fresh produce of immuno-PCR detection of Escherichia coli O157:H7. FEMS Immunol. Med. Microbiol. 23(3), 213–220 (1999)
L.C. Shriver-Lake, Y.S. Shubin, F.S. Ligler, Detection of staphylococcal enterotoxin B in spiked food samples. J. Food Prot. 66(10), 1851–1856 (2003)
S.M. Henning, W. Aronson, Y. Niu, F. Conde, N.H. Lee, N.P. Seeram, R.P. Lee, J. Lu, D.M. Harris, A. Moro, J. Hong, L. Pak-Shan, R.J. Barnard, H.G. Ziaee, G. Csathy, V.L. Go, H. Wang, D. Heber, Tea polyphenols and theaflavins are present in prostate tissue of humans and mice after green and black tea consumption. J. Nutr. 136(7), 1839–1843 (2006)
J. Homola, J. Dostalek, S. Chen, A. Rasooly, S. Jiang, S.S. Yee, Spectral surface plasmon resonance biosensor for detection of staphylococcal enterotoxin B in milk. Int. J. Food Microbiol. 75(1–2), 61–69 (2002)
J.R. Son, G. Kim, A. Kothapalli, M.T. Morgan, D. Ess, Detection of Salmonella enteritidis using a miniature optical surface plasmon resonance biosensor. J. Phys. 61, 1086–1090 (2007)
K.E. Sapsford, C.R. Taitt, N. Loo, F.S. Ligler, Biosensor detection of botulinum toxoid A and staphylococcal enterotoxin B in food. Appl. Environ. Microbiol. 71(9), 5590–5592 (2005)
G.P. Anderson, J.L. Liu, M.L. Hale, R.D. Bernstein, M. Moore, M.D. Swain, E.R. Goldman, Development of anti-ricin single domain antibodies toward detection and therapeutic reagents. Anal. Chem. 80(24), 9604–9611 (2008)
S. Bhaduri, B. Cottrell, Sample preparation methods for PCR detection of Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes on beef chuck shoulder using a single enrichment medium. Mol. Cell. Probes 15(5), 267–274 (2001)
A.A. Bhagwat, Simultaneous detection of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella strains by real-time PCR. Int. J. Food Microbiol. 84(2), 217–224 (2003)
A.M. Valadez, C.A. Lana, S.I. Tu, M.T. Morgan, A.K. Bhunia, Evanescent wave fiber optic biosensor for Salmonella detection in food. Sensors 9(7), 5810–5824 (2009)
D.V. Lim, Detection of microorganisms and toxins with evanescent wave fiber-optic biosensors. Proc. IEEE 91(6), 902–907 (2003)