Displacement immunoassay for the detection of ochratoxin A using ochratoxin B modified glass beads

Abstract

We report here the development of a new assay for the detection of ochratoxin A (OTA) based on the use of its dechlorinated analogue, ochratoxin B (OTB), in a displacement immunoassay. OTB was immobilised on controlled-pore glass beads followed by the binding of anti-OTA antibody, with anti-IgG antibody peroxidase conjugate used as a label. In this manner, an original bio-sensing material was obtained. Upon incubation of this material with OTA, the toxin competes with OTB for the binding sites of the anti-OTA antibodies and releases the antibody-tagged peroxidase complex into the solution. Compared to classic competitive immunoassays, this newly developed displacement immunoassay presents a similar detection limit and assay time. Moreover, the detection, based on the activity of the horseradish peroxidase, is performed for the first time in situ using wine samples.

Steps in the construction of the OTB based bio-sensing glass material (1–5) and the displacement reaction (step 6) upon incubation with ochratoxin A

This is a preview of subscription content, access via your institution.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

References

  1. 1.

    Gekle M, Silbernagl S (1993) Mechanism of ochratoxin A-induced reduction of glomerular filtration rate in rats. J Pharmacol Exp Ther 267(1):316–321

    CAS  Google Scholar 

  2. 2.

    Shlosberg A, Elkin N, Malkinson M, Orgad U, Hanji V, Bogin E, Weisman Y, Meroz M, Bock R (1997) Severe hepatopathy in geese and broilers associated with ochratoxin in their feed. Mycopathologia 138(2):71–76

    Article  CAS  Google Scholar 

  3. 3.

    Muller G, Kielstein P, Rosner H, Berndt A, Heller M, Kohler H (1999) Studies of the influence of ochratoxin A on immune and defence reactions in weaners. Mycoses 42(7–8):495–505

    Article  CAS  Google Scholar 

  4. 4.

    Cavin C, Delatour T, Marin-Kuan M, Fenaille F, Holzhauser D, Guignard G, Bezencon C, Piguet D, Parisod V, Richoz-Payot J, Schilter B (2009) Ochratoxin A—mediated DNA and protein damage: Roles of nitrosative and oxidative stresses. Toxicol Sci 110(1):84–94

    Article  CAS  Google Scholar 

  5. 5.

    Mantle PG, Faucet-Marquis V, Manderville RA, Squillaci B, Pfohl-Leszkowicz A (2010) Structures of covalent adducts between DNA and ochratoxin a: a new factor in debate about genotoxicity and human risk assessment. Chem Res Toxicol 23(1):89–98

    Article  CAS  Google Scholar 

  6. 6.

    Tatu CA, Orem WH, Finkelman RB, Feder GL (1998) The etiology of Balkan endemic nephropathy: still more questions than answers. Environ Health Perspect 106(11):689–700

    Article  CAS  Google Scholar 

  7. 7.

    Visconti A, Pascale M, Centonze G (2001) Determination of ochratoxin A in wine and beer by immunoaffinity column cleanup and liquid chromatographic analysis with fluorometric detection: collaborative study. J AOAC Int 84:1818–1827

    CAS  Google Scholar 

  8. 8.

    Rusanova TY, Beloglazova NV, Goryacheva IY, Lobeau M, Peteghem C, Saeger SD (2009) Non-instrumental immunochemical tests for rapid ochratoxin A detection in red wine. Anal Chim Acta 653:97–102

    Article  CAS  Google Scholar 

  9. 9.

    Radoi A, Dumitru L, Barthelmebs L, Marty JL (2009) Ochratoxin A in some french wine: application of a direct competitive ELISA based on an OTA-HRP conjugate. Anal Lett 42:1187–1202

    Article  CAS  Google Scholar 

  10. 10.

    Prieto-Simon B, Campas M, Marty JL, Noguer T (2008) Novel highly-performing immunosensor-based strategy for ochratoxin A detection in wine samples. Biosens Bioelectron 23(7):995–1002

    Article  CAS  Google Scholar 

  11. 11.

    Vidal JC, Duato P, Bonel L, Castillo JR (2009) Use of polyclonal antibodies to ochratoxin A with a quartz-crystal microbalance for developing real-time mycotoxin piezoelectric immunosensors. Anal Bioanal Chem 394(2):575–582

    Article  CAS  Google Scholar 

  12. 12.

    Holt DB, Kusterbeck AW, Ligler FS (2000) Continuous flow displacement immunosensors: a computational study. Anal Biochem 287:234–242

    Article  CAS  Google Scholar 

  13. 13.

    Whelan JP, Kusterbeck AW, Wemhoff GA, Bredehorst R, Ligler F (1993) Continuous-flow immunosensor for detection of explosives. Anal Chem 65:3561–3565

    Article  CAS  Google Scholar 

  14. 14.

    Green TM, Charles PT, Anderson GP (2002) Detection of 2,4,6-trinitrotoluene in seawater using a reversed-displacement immunosensor. Anal Biochem 310:36–41

    Article  CAS  Google Scholar 

  15. 15.

    Kaptein WA, Zwaagstra JJ, Venema K, Ruiters M, Korf J (1997) Analysis of cortisol with a flow displacement immunoassay. Sens Act B 45:63–69

    Article  Google Scholar 

  16. 16.

    Voort D, Pelsers M, Korf J, Hermens WT, Glatz JFC (2003) Development of a displacement immunoassay for human heart-type fatty acid-binding protein in plasma: the basic conditions. Biosens Bioelectron 19:465–471

    Article  Google Scholar 

  17. 17.

    Acharya D, Dhar TK (2008) A novel broad-specific noncompetitive immunoassay and its application in the determination of total aflatoxins. Anal Chim Acta 630(1):82–90

    CAS  Google Scholar 

  18. 18.

    Harris JP, Mantle PG (2001) Biosynthesis of ochratoxins by Aspergillus ochraceus. Phytochem 58(5):709–716

    Article  CAS  Google Scholar 

  19. 19.

    Dietrich DR, O’Brien EO, Stack ME, Heussner AH (2001) Species- and sex-specific renal cytotoxicity of ochratoxin A and B in vitro. Exp Toxic Pathol 53(2–3):215–225

    Article  CAS  Google Scholar 

  20. 20.

    Bonel L, Vidal JC, Duato P, Castillo JR (2011) An electrochemical competitive biosensor for ochratoxin A based on a DNA biotinylated aptamer. Biosens Bioelectron 26(7):3254–3259

    Article  CAS  Google Scholar 

  21. 21.

    Sheng L, Ren J, Miao Y, Wang J, Wang E (2011) PVP-coated graphene oxide for selective determination of ochratoxin A via quenching fluorescence of free aptamer. Biosens Bioelectron In Press, Corrected Proof

  22. 22.

    Chung SWC, Kwong KP (2007) Determination of ochratoxin A at parts-per-trillion levels in cereal products by immunoaffinity column cleanup and high-performance liquid chromatography/mass spectrometry. J AOAC Int 90(3):773–777

    CAS  Google Scholar 

  23. 23.

    Remiro R, Ibanez-Vea M, Gonzalez-Penas E, Lizarraga E (2010) Validation of a liquid chromatography method for the simultaneous quantification of ochratoxin A and its analogues in red wines. J Chromat A 1217:8249–8256

    Article  CAS  Google Scholar 

  24. 24.

    Han Z, Zheng Y, Luan L, Ren Y, Wu Y (2010) Analysis of ochratoxin A and ochratoxin B in traditional Chinese medicines by ultra-high-performance liquid chromatography-tandem mass spectrometry using [13C20]-ochratoxin A as an internal standard. J Chromat A 1217(26):4365–4374

    Article  CAS  Google Scholar 

  25. 25.

    Lates V, Popescu IC, Marty JL (2011) Determination of antioxidant capacity by using xanthine oxidase bioreactor coupled with flow-through H2O2 amperometric biosensor. Electroanal 23:728–736

    CAS  Google Scholar 

  26. 26.

    Kahraman MV, Bayramoglu G, Kayaman-Apohan N, Güngör A (2007) alpha-Amylase immobilization on functionalized glass beads by covalent attachment. Food Chem 104(4):1385–1392

    Article  CAS  Google Scholar 

  27. 27.

    Nakamura H, Murakami Y, Yokoyama K, Tamiya E, Karube I, Suda M, Uchiyama S (2001) A compactly integrated flow cell with a chemiluminescent FIA system for determining lactate concentration in serum. Anal Chem 73:373–378

    Article  CAS  Google Scholar 

  28. 28.

    Molina L, Messina G, Stege P, Salinas E, Raba J (2008) Immuno-column for on-line quantification of human serum IgG antibodies to Helicobacter pylori in human serum samples. Talanta 76:1077–1082

    Article  CAS  Google Scholar 

  29. 29.

    Krylov AV, Adamzig H, Walter AD, Lochel B, Kurth E, Pulz O, Szeponik J, Wegerich F, Lisdat F (2006) Parallel generation and detection of superoxide and hydrogen peroxide in a fluidic chip. Sens Actuators B 119(1):118–126

    Article  Google Scholar 

  30. 30.

    Rabbany SY, Piervincenzi R, Judd L, Kusterbeck AW, Bredehorst R, Hakansson K, Ligler F (1997) Assessment of heterogeneity in antibody–antigen displacement reactions. Anal Chem 69:175–182

    Article  CAS  Google Scholar 

  31. 31.

    Fintschenko Y, Wilson G (1998) Flow injection immunoassays: a review. Mikrochim Acta 129:7–18

    Article  CAS  Google Scholar 

  32. 32.

    Hock B (2000) Immunoassays. In: Bilitewski U, Turner APF (eds) Biosensors for environmental monitoring. Harwood, Amsterdam, p 105

    Google Scholar 

  33. 33.

    Valero A, Marin S, Ramos AJ, Sanchis V (2007) Survey: ochratoxin A in European special wines. Food Chem 108:593–599

    Article  Google Scholar 

  34. 34.

    Sibanda L, Saeger SD, Peteghem C (2002) Optimization of solid-phase clean-up prior to liquid chromatographic analysis of ochratoxin A in roasted coffee. J Chromat A 959:327–330

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors gratefully acknowledge the “Agence Universitaire de la Francophonie” (AUF) for funding Vasilica Lates.

Author information

Affiliations

Authors

Corresponding author

Correspondence to Jean-Louis Marty.

Electronic supplementary material

Below is the link to the electronic supplementary material.

ESM 1

(PDF 37.2 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Lates, V., Yang, C., Popescu, I.C. et al. Displacement immunoassay for the detection of ochratoxin A using ochratoxin B modified glass beads. Anal Bioanal Chem 402, 2861–2870 (2012). https://doi.org/10.1007/s00216-012-5721-4

Download citation

Keywords

  • Ochratoxin B
  • Displacement immunoassay
  • Controlled-pore glass
  • Ochratoxin A