Development of a QCM (Quartz Crystal Microbalance) Biosensor to the Detection of Aflatoxin B1


In this study, we have used a direct immunoassay where the simple binding between antigen and an antibody is detected. Immunoassays were performed in a drop system, monitoring the frequency decrease of the quartz-crystal microbalance device because of mass increasing during immunoreaction. The QCM sensor was coated on both sides by gold electrodes, only one side of the crystal (liquid side) was in contact with the solution; the other side (contact side) was always dry. We tested a piezoelectric immunosensor for aflatoxin B1 (AFLA-B1) mycotoxin detection through the immo- bilization of DSP-anti-AFLAB1 antibody (AFLA-B1-Ab anti AFLAB1) on gold-coated quartz crystals (AT-cut/5 MHz). The DSP (3,3’-Dithiodipropionic-acid-di-N-hydroxysuccinimide ester) was used for the covalent attachment of the proteins. The piezoelectric crystal electrodes were pretreated by DSP for 15 min, rinsed with water and dried in a gentle flow of nitrogen gas. Then the DSP-coated crystals were installed in a sample holder and exposed to the anti-AFLAB1 antibody and to the AFLA-BI. Frequency and resistance shifts (Δf and ΔR) were measured simultaneously. Δf versus AFLA-BI concentrations in the range of 0.5 - 10 ppb exhibited a perfect linear correlation with a coefficient of above 0.998.

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Spinella, K. , Mosiello, L. , Palleschi, G. and Vitali, F. (2013) Development of a QCM (Quartz Crystal Microbalance) Biosensor to the Detection of Aflatoxin B1. Open Journal of Applied Biosensor, 2, 112-119. doi: 10.4236/ojab.2013.24015.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] R. J. Cole and R. H. Cox, “Handbook of Toxic Fungal Metabolites,” Academic Press, New York, 1981.
[2] International Agency for Research on Cancer, “IARC Monographs on the Evaluations of Carcinogenic Risks to Humans,” IARC, Lyon, Vol. 56, 1993, pp. 489-521.
[3] O. M. Moss, “Risk Assessment for Aflatoxins in Food-Stuffs,” International Biodeterioration & Biodegradation, Vol. 50, No. 3-4, 2002, pp. 137-142.
[4] J. Yang, F. Bai, K. Zhang, S. Bai, X. Peng, X. Ding, Y. Li, J. Zhang and L. Zhao, “Effects of Feeding Corn Naturally Contaminated with Aflatoxin B1 and B2 on Hepatic Functions of Broilers,” Poultry Science, Vol. 91, No. 11, 2012, pp. 2792-2801.
[5] M. Miraglia, C. Brera and M. Colatosti, “Application of Biomarkers to Assessment of Risk to Human Health from Exposure to Mycotoxins,” Microchemical Journal, Vol. 54, No. 4, 1996, pp. 472-477.
[6] A. De Girolamo, M. McKeague, D. J. Miller, M. C. DeRosa and A. Visconti, “Determination of Ochratoxin A in Wheat after Clean-Up through a DNA Aptamer-Based Solid Phase Extraction Column,” Food Chemistry, Vol. 127, No. 3, 2011, pp. 1378-1384.
[7] H. P. Van Egmond and A. R. O. Jonker, “Worldwide Regulations for Mycotoxins in Food and Feed in 2003,” FAO Food and Nutrition Paper 81, Report of the Food and Agriculture Organization of the United Nations, Rome, 2004.
[8] A. Fernandez, R. Belio, J. J. Ramos, M. C. Sanz and T. Saez, “Aflatoxins and Their Metabolites in the Tissues, Faeces and Urine from Lambs Feeding on an Aflatoxin-Contaminated Diet,” Journal of the Science of Food and Agriculture, Vol. 74, No. 2, 1997, pp. 161-168.<161::AID-JSFA783>3.0.CO;2-D
[9] J. Jaimez, C. A. Fente, B. I. Vazquez, C. M. Franco, A. Capeda, G. Mahhuzier and P. Prognon, “Application of the Assay of Aflatoxins by Liquid Chromatography with Fluorescence Detection in Food Analysis,” Journal of Chromatography A, Vol. 882, No. 1-2, 2000, pp. 1-10.
[10] S. Piermarini, L. Micheli, N. H. S. Ammida, G. Palleschi and D. Moscone, “Electrochemical Immunosensor Array Using a 69-Well Screen-Printed Microplate for Aflatoxin B1 Detection,” Biosensors & Bioelectronics, Vol. 22, No. 7, 2007, pp. 1434-1440.
[11] J. Stroka and E. Anklam, “New Strategies for the Screening and Determination of Aflatoxins and the Detection of Aflatoxin-Producing Moulds in Food and Feed,” TrAC Trends in Analytical Chemistry, Vol. 21, No. 2, 2002, pp. 90-95.
[12] S. Piermarini, G. Volpe, F. Ricci, L. Micheli, D. Moscone, G. Palleschi, M. Fuhrer, R. Krska and S. Baumgartner, “Rapid Screening Electrochemical Methods for Aflatoxin B1 and Type-A Trichothecenes: A Preliminary Study,” Analytical Letters, Vol. 40, No. 7, 2007, pp. 1333-1346.
[13] L. Micheli, R. Grecco, M. Badea, D. Moscone and G. Palleschi, “An Electrochemical Immunosensor for Aflatoxin M1 Determination in Milk Using Screen Printed Electrodes,” Biosensor and Bioelectronics, Vol. 21, 2006, pp. 1424-1433.
[14] T. K. Dutta and P. Das, “Isolation of Aflatoxigenic Strains of Aspergillus and Detection of Aflatoxin B1 from Feeds in India,” Mycopathologia, Vol. 151, No. 1, 2000, pp. 29- 33.
[15] H. Ayciek, A. Aksoy and S. Saygi, “Determination of Aflatoxin Levels in Some Dairy and Food Products Consumed in Ankara, Turkey,” Food Control, Vol. 16, No. 3, 2005, pp. 263-266.
[16] H. Zheng, C. W. Humphney, R. S. King and J. L. Richard, “A Review of Rapid Methods for the Analysis of Aflatoxins,” Mycopathologia, Vol. 159, 2005, pp. 1-9.
[17] T. Yaroglu, H. H. Oruc and M. Tayar, “Aflatoxin M1 Levels in Cheese Samples from Some Provinces of Turkey,” Food Control, Vol. 16, No. 10, 2005, pp. 883-885.
[18] A. Y. Kolosova, W. B. Shim, Z. Y. Yang, S. A. Eremin and D. H. Chung, “Direct Competitive ELISA Based on a Monoclonal Antibody for Detection of Aflatoxin B1. Stabilization of ELISA Kit Components and Application to Grain Samples,” Analytical and Bioanalytical Chemistry, Vol. 384, No. 1, 2006, pp. 286-294.
[19] S. Rastogi, P. D. Divedi, S. K.. Khanna and M. Das, “Detection of Aflatoxin M1 Contamination in Milk and Infant Milk Products from Indian Markets by ELISA,” Food Control, Vol. 15, 2004, pp. 287-290.
[20] R. M. Pemberton, R. Pittson, N. Biddle, G. A. Drago and J. P. Hart, “Studies towards the Development of a Screen-Printed Carbon Electrochemical Immunosensor Array for Mycotoxins: A Sensor for Aflatoxin B1,” Analytical Letters, Vol. 39, No. 8, 2006, pp. 1573-1586.
[21] J. H. O. Owino, A. Ignaszak, A. Al-Ahmed, P. G. L. Baker, H. Alemu, J. C. Ngila and E. I. Iwuoha, “Modelling of the Impedimetric Responses of an Aflatoxin B1 Immunosensor Prepared on an Electrosynthetic Polyaniline Platform,” Analytical and Bioanalytical Chemistry, Vol. 388, No. 5-6, 2007, pp. 1069-1074.
[22] N. Adanyi, I. A. Levkovets, S. Rodriguez-Gil, A. Ronald, M. Varadi and I. Szendro, “Development of Immunosensor Based on OWLS Technique for determining Aflatoxin B1 and Ochratoxin A,” Biosensor and Bioelectronics, Vol. 22, No. 6, 2007, pp. 797-802.
[23] A. Sun, Q. Qi, Z. L. Dong and K. Z. Liang, “An Electro-chemical Enzyme Immunoassay for Aflatoxin B1 Based on Bio-Electrocatalytic Reaction with Room-Temperature Ionic Liquid and Nanoparticle Modified Electrodes,” Sensing and Instrumentation for Food Quality and Safety, Vol. 2, No. 1, 2008, pp. 43-50.
[24] J. Ren, F. He, S. Yi and X. Cui, “A New MSPQC for Rapid Growth and Detection of Mycobacterium tuberculosis,” Biosensors and Bioelectronics, Vol. 24, No. 3, 2008, pp. 403-409.
[25] X. Tu, Q. Xie, S. Jiang and S. Yao, “Electrochemical Quartz Crystal Impedance Study on the Overoxidation of Polypyrrole-Carbon Nanotubes Composite Film for Amperometric Detection of Dopamine,” Biosensors and Bio-electronics, Vol. 22, No. 12, 2007, pp. 2819-2826.
[26] S. F. Chou, W. L. J. Hsu, M. Hwang and C. Y. Chen, “Determination of Alpha-Fetoprotein in Human Serum by a Quartz Crystal Microbalance-Based Immunosensor,” Clinical Chemistry, Vol. 48, 2002, pp. 913-918.
[27] B. Zhang, Q. Mao, X. Zhang, T. Jiang, M. Chen, F. Yu and W. Fu, “A Novel Piezoelectric Quartz Micro-Array Immunosensor Based on Self-Assembled Monolayer for Determination of Human Chorionic Gonadotropin,” Biosensors and Bioelectronics, Vol. 15, 2004, pp. 711-720.
[28] G. Z. Sauerbrey, “Use of Quartz Vibration for Weighing Thin Films on a Microbalance,” Journal of Physik, Vol. 155, 1959, pp. 206-212.

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