QCM‐Based Immunosensor for the Determination of Ochratoxin A

Abstract A piezoelectric immunosensor based on a competitive format was developed for determination of ochratoxin A (OTA) concentration. Surface modifications via two self‐assembled monolayers (SAMs) were investigated respectively and a better result was obtained with the SAM of 16‐mercaptohexadecanoic acid (16‐MHDA). The quartz crystal microbalance (QCM)‐based immunosensor was fabricated by immobilizing anti‐OTA antibodies onto the surface of the 16‐MHDA‐modified electrode, and allowing competition between free OTA and that conjugated with BSA to occur. The assay exhibited a working range of 50–1000 ng/mL and a detection limit of 16.1 ng/mL. Studies of interference and matrix effects were performed to evaluate the feasibility of the developed immunosensor for the direct analysis of OTA in real samples. Recoveries were conducted at 50, 200, and 1000 ng/g and were determined to be in the range of 142%–76%. The OTA assay is specific. No cross‐reactivates were observed with citrinin.

[1]  A. Turner,et al.  Immunosensor for okadaic acid using quartz crystal microbalance , 2002 .

[2]  A. Suleiman,et al.  A Piezoelectric Immunosensor for the Detection of Cocaine , 1996 .

[3]  Y. Ueno,et al.  Enzyme‐linked immunosorbent assay for detection and survey of ochratoxin a in livestock sera and mixed feeds , 1990 .

[4]  Kenji Yokoyama,et al.  Highly sensitive quartz crystal immunosensors for multisample detection of herbicides , 1995 .

[5]  E. M. de Groene,et al.  Mutagenicity and genotoxicity of the mycotoxin ochratoxin A. , 1996, Environmental toxicology and pharmacology.

[6]  P. Skládal,et al.  Construction and characterization of the direct piezoelectric immunosensor for atrazine operating in solution. , 1997, Biosensors & bioelectronics.

[7]  J. Pitt,et al.  Fungi and Food Spoilage , 1987 .

[8]  D. Chan,et al.  Immunosensors--principles and applications to clinical chemistry. , 2001, Clinica chimica acta; international journal of clinical chemistry.

[9]  O. Busto,et al.  Solid-phase extraction applied to the determination of ochratoxin A in wines by reversed-phase high-performance liquid chromatography. , 2000, Journal of chromatography. A.

[10]  T. Troxell,et al.  Identification and reduction of sources of dietary lead in the United States. , 1996, Food additives and contaminants.

[11]  S. Babacan,et al.  Evaluation of antibody immobilization methods for piezoelectric biosensor application. , 2000, Biosensors & bioelectronics.

[12]  Fook Tim Chew,et al.  Design and Application of Piezoelectric Quartz Crystal-based Immunoassay , 2000 .

[13]  C. Nakamura,et al.  Conventional detection of 2,4-dinitrophenol using quartz crystal microbalance , 2003, IEEE Transactions on Ultrasonics, Ferroelectrics and Frequency Control.

[14]  John G. Anderson,et al.  Role of mycotoxins in human and animal nutrition and health. , 1995, Natural toxins.

[15]  Capture agents for a quartz crystal microbalance-continuous flow biosensor: functionalized self-assembled monolayers on gold , 1999 .

[16]  Jian Zhang,et al.  Comparison of surface plasmon resonance spectroscopy and quartz crystal microbalance for human IgE quantification , 2004 .

[17]  Heidi R. C. Dietrich,et al.  Biosensors and multiple mycotoxin analysis , 2003 .

[18]  A. Cepeda,et al.  Simultaneous high-performance liquid chromatographic determination of ochratoxin A and citrinin in cheese by time-resolved luminescence using terbium , 1996 .

[19]  A. Leitner,et al.  Comparison of methods for the determination of ochratoxin A in wine , 2002 .

[20]  F. Pariente,et al.  A quartz crystal microbalance assay for detection of antibodies against the recombinant African swine fever virus attachment protein p12 in swine serum , 1998 .

[21]  H. Reinhard,et al.  Reversed-phase liquid chromatographic behavior of the mycotoxins citrinin and ochratoxin A. , 1999, Journal of chromatography. A.

[22]  J. Chiba,et al.  A sensitive enzyme-linked immunosorbent assay of ochratoxin A based on monoclonal antibodies. , 1989, Toxicon : official journal of the International Society on Toxinology.

[23]  S. Patel,et al.  Survey of ochratoxin A in UK retail coffees. , 1997, Food additives and contaminants.

[24]  G. E. Who,et al.  IARC monographs on the evaluation of carcinogenic risks to humans. v. 51: Coffee, tea, mate, methylxanthines and methylglyoxal , 1991 .

[25]  Giuseppe Palleschi,et al.  Electrochemical immunosensor for determination of aflatoxin B1 in barley , 2004 .

[26]  P. Mantle,et al.  Ochratoxin formation in Aspergillus ochraceus with particular reference to spoilage of coffee. , 2000, International journal of food microbiology.

[27]  P. Scott,et al.  Risk assessment of the mycotoxin ochratoxin A. , 1989, Biomedical and environmental sciences : BES.

[28]  P. Scott Recent developments in methods of analysis for mycotoxins in foodstuffs , 1993 .

[29]  C. V. Van Peteghem,et al.  Optimization of solid-phase clean-up prior to liquid chromatographic analysis of ochratoxin A in roasted coffee. , 2002, Journal of chromatography. A.

[30]  S. Drost,et al.  Characterization of immobilization methods for African swine fever virus protein and antibodies with a piezoelectric immunosensor. , 1998, Biosensors & bioelectronics.

[31]  W. Tsai,et al.  Development of a piezoelectric immunosensor for the detection of alpha-fetoprotein , 2005 .

[32]  M. Mascini,et al.  Surface modifications for the development of piezoimmunosensors. , 1998, Biosensors & bioelectronics.

[33]  R. R. Marquardt,et al.  A review of recent advances in understanding ochratoxicosis. , 1992, Journal of animal science.

[34]  Liberty Sibanda,et al.  A collaborative study to validate novel field immunoassay kits for rapid mycotoxin detection. , 2002, International journal of food microbiology.