A Sensitive Chemiluminescent Assay for Aflatoxin B1
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Abstract
In this study a chemiluminescent label was substituted for an enzyme marker used in AFB1 ELISA and the minimum detection limits of the direct/modified indirect competitive chemiluminescence immunoassay (CIA), the direct competitive CIA (2 step method), the direct competitive CIA (1 step, 30 min method), and the direct competitive CIA (1 step, 5 min method) were shown to be 10.0, 5.1, 5.6, and 6.8 pg ml-1 respectively. In a direct comparison between different labeling methods but using the same antibodies, the direct competitive CIA (1 step, 5 min method) is superior in sensitivity to the AFB1 ELISA (detection limit 200 pg ml-1). In addition to an increase in sensitivity, the assay time was also reduced. The sensitivity of the assay was not affected by 6.25% (v/v) methanol and required a 1:8 dilution of the peanut extract [using 80% (v/v) methanol as extracting solvent] was required to avoid matrix interference.
Keywords: aflatoxin B1, chemiluminescent label, enzyme marker, immunoassay
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References
[2] International Agency for Research on Cancer Monographs. Evaluation of carcinogenic risk of chemicals to humans, IARC Monograph Supp, 7, 1987, 83-87.
[3] Food and Agriculture Organization/World Health Organization, Annex 1-Aflatoxins: Summary and Conclusions, The Joint FAO/WHO Expert Committee on Food Additives, 49th meeting, 1997, 16-17.
[4] Stenman, U.H. Quantitation of antigen. In: Herzenberg, L.A. Weir, D.M. Herzenberg, L.A. and Blackwell, C. Weir’s handbook of experimental immunology. 5 th ed. Massachusetts, Blackwell Science, 1996, pp. 45.1-45.7.
[5] Ekins, R.P. and Chu, F.W. Multianalyte microspot immunoassay: microanalytical “compact disk” of the future, Clinical Chemistry, 37, 1991, 1955-1967.
[6] Alkan, S. Akdis, C. and Towbin, H. Chemiluminescent and enzyme-linked immunoassays foe sensitive detection of human IFN-γ, Journal of Immunoassay, 15, 1994, 217-238.
[7] Dudley, R.S. Chemiluminescence immunoassay: An alternative to RIA, Laboratory Medicine, 21, 1990, 216-22.
[8] Rongen, H.A.H. Hoetelmans, R.M.W. Bult, A. and Van Bennekom, W.P. Chemiluminescence and immunoassays, Journal of Pharmacological and Biomedical Analysis, 12, 1994, 433-462.
[9] Candlish, A.A.G. The development of monoclonal antibody based enzyme immunoassays for ochratoxin A and aflatoxin B1. Glasgow, University of Strathclyde, 1987.
[10] Candlish, A.A.G. Stimson, W.H. and Smith, J.E. A monoclonal antibody to aflatoxin B1: Detection of the mycotoxin by enzyme immunoassay, Letters in Applied Microbiology, 1, 1985, 57-61.
[11] Goodbrand, I.A. Stimson, W.H. and Smith, J.E. A monoclonal antibody to T-2 toxin, Letters in Applied Microbiology, 5, 1987, 97-99.
[12] Chu, F.S. Development of antibodies against aflatoxins. In: Eaton, D.L. and Groopman, J.D. The toxicology of aflatoxins. New York, Academic Press, 1994, pp. 451-490.
[13] Fan, T.S.L. and Chu, F.S. An indirect ELISA for detection of aflatoxin B1 in corn and peanut butter, Journal of Food Protection, 47, 1984, 263-266.
[14] Richardson, A.P. Kim, J.B. Barnard, G.J. Collins, W.P. and McCapra, F. Chemiluminescence immunoassay of plasma progesterone, with progesterone-acridinium ester used as the labeled antigen, Clinical Chemistry, 31, 1985, 1664-1668.
[15] Kawamura, O. Nagayama, S. Sato, S. Ohtani, K. Ueno, I. and Ueno, Y. A monoclonal antibody based enzyme-linked immunosorbent assay for aflatoxin B1 in peanut products, Mycotoxin Research, 4, 1988, 75-88.
[16] Swinscow, T.D.V. In: British Medical Association. Statistics at square one. 8 th ed. London, Dawson and Global, 1983, pp. 21-28.
[17] Whitaker, T.B. Dickens, J.W. and Giesbrecht, F.G. Effect of methanol concentration and solvent: peanut ratio on extraction of aflatoxin from raw peanut, Journal of the Association of Official analytical Chemists, 67, 1984, 35-36.
[18] Whitaker, T.B. and Dickens, J.W. Optimal methanol concentration and solvent/peanut ratio for extraction of aflatoxin from raw peanuts by modified AOAC method II, Journal of the Association of Official Analytical Chemists, 69, 1986, 508-10.
[19] El-Nakib. Pestka, J.J. and Chu, F.S. Determination of aflatoxin B1 in corn, wheat, and peanut butter by enzyme-linked immunosorbent assay and solid phase radioimmunoassay, Journal of the Association of Official Analytical Chemists, 64, 1981, 1077-1082.
[20] Ram, B.P. Hart, L.P. Shotwell, O.I. and Pestka, J.J. Enzyme-linked immunosorbent assay for aflatoxin B1 in naturally contaminated corn and cottonseed, Journal of the Association of Official Analytical Chemists, 69, 1986, 904-907.
[21] Chu, F.S. Fan, T.S.L. Zhang, G-S. Xu, Y.C. Faust, S. and McMahon, P.L. Improved enzyme-linked immunosorbent assay for AFB1 in agricultural commodities, Journal of the Association of Official Analytical Chemists, 70, 1987, 854-857.