Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1996 Jun;62(6):1880–1884. doi: 10.1128/aem.62.6.1880-1884.1996

Dipstick enzyme immunoassay to detect Fusarium T-2 toxin in wheat.

S De Saeger 1, C Van Peteghem 1
PMCID: PMC167966  PMID: 8787386

Abstract

A dipstick enzyme immunoassay for the rapid detection of Fusarium T-2 toxin in wheat was developed. An Immunodyne ABC membrane was precoated with rabbit anti-mouse immunoglobulins. After the strips were immersed in a solution of monoclonal anti-T-2 toxin antibodies, a direct competitive enzyme immunoassay was performed. This assay included the incubation of the antibody-coated dipsticks in a mixture of sample and T-2 toxin-horseradish peroxidase conjugate. Afterwards, the strips were placed in a chromogen-containing substrate solution (H202-3,3',5,5'-tetramethylbenzidine) for color reaction. The dot color intensity of toxin-positive dipsticks was visually distinguishable from that of the negative control. A portable colorimeter was used to confirm and quantify the visual observations. With coated strips, the tests could be performed in 45 min. The visual detection limit for T-2 toxin in buffer solution was 0.25 ng/ml. Artificially infected wheat samples were extracted with 80% methanol-water. A dilution of the raw extract of 1:8 was sufficient to avoid matrix effects. It was possible to make visually a clear distinction between the negative control and a wheat extract spiked with 12 ng/g.

Full Text

The Full Text of this article is available as a PDF (226.0 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abouzied M. M., Pestka J. J. Simultaneous screening of fumonisin B1, aflatoxin B1, and zearalenone by line immunoblot: a computer-assisted multianalyte assay system. J AOAC Int. 1994 Mar-Apr;77(2):495–501. [PubMed] [Google Scholar]
  2. Barna-Vetró I., Gyöngyösi A., Solti L. Monoclonal antibody-based enzyme-linked immunosorbent assay of Fusarium T-2 and zearalenone toxins in cereals. Appl Environ Microbiol. 1994 Feb;60(2):729–731. doi: 10.1128/aem.60.2.729-731.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Butler J. E., Ni L., Brown W. R., Joshi K. S., Chang J., Rosenberg B., Voss E. W., Jr The immunochemistry of sandwich ELISAs--VI. Greater than 90% of monoclonal and 75% of polyclonal anti-fluorescyl capture antibodies (CAbs) are denatured by passive adsorption. Mol Immunol. 1993 Sep;30(13):1165–1175. doi: 10.1016/0161-5890(93)90135-x. [DOI] [PubMed] [Google Scholar]
  4. Gendloff E. H., Pestka J. J., Swanson S. P., Hart L. P. Detection of T-2 toxin in Fusarium sporotrichioides-infected corn by enzyme-linked immunosorbent assay. Appl Environ Microbiol. 1984 May;47(5):1161–1163. doi: 10.1128/aem.47.5.1161-1163.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kim M. S., Doyle M. P. Dipstick immunoassay to detect enterohemorrhagic Escherichia coli O157:H7 in retail ground beef. Appl Environ Microbiol. 1992 May;58(5):1764–1767. doi: 10.1128/aem.58.5.1764-1767.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Laamanen I., Veijalainen P. Factors affecting the results of T-2 mycotoxin ELISA assay. Food Addit Contam. 1992 Jul-Aug;9(4):337–343. doi: 10.1080/02652039209374079. [DOI] [PubMed] [Google Scholar]
  7. Lacey J., Ramakrishna N., Candlish A. A., Smith J. E. Immunoassay of ochratoxin and other mycotoxins from a single extract of cereal grains utilizing monoclonal antibodies. IARC Sci Publ. 1991;(115):97–103. [PubMed] [Google Scholar]
  8. May M. E., Vermeulen C. W. Enhanced enzyme-linked immunosorbant assay on membranes for the identification of mutants and pathogens. Life Sci. 1991;49(12):865–868. doi: 10.1016/0024-3205(91)90171-7. [DOI] [PubMed] [Google Scholar]
  9. Morissette C., Goulet J., Lamoureux G. Rapid and sensitive sandwich enzyme-linked immunosorbent assay for detection of staphylococcal enterotoxin B in cheese. Appl Environ Microbiol. 1991 Mar;57(3):836–842. doi: 10.1128/aem.57.3.836-842.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Pfund W. P., Bourdage J. S. The conformation-sensitive immunoassay: a membrane based ELISA system for identifying antibodies sensitive to alterations of protein conformation. Mol Immunol. 1990 Jun;27(6):495–502. doi: 10.1016/0161-5890(90)90068-b. [DOI] [PubMed] [Google Scholar]
  11. Ramadass P., Meerarani S., Padmanaban V. D. Dipstick enzyme immunoassay for rinderpest antibody in cattle. Vet Microbiol. 1993 Sep;36(3-4):385–388. doi: 10.1016/0378-1135(93)90105-g. [DOI] [PubMed] [Google Scholar]
  12. Ramakrishna N., Lacey J., Candlish A. A., Smith J. E., Goodbrand I. A. Monoclonal antibody-based enzyme linked immunosorbent assay of aflatoxin B1, T-2 toxin, and ochratoxin A in barley. J Assoc Off Anal Chem. 1990 Jan-Feb;73(1):71–76. [PubMed] [Google Scholar]
  13. Savage H. M., Duncan J. F., Roberts D. R., Sholdt L. L. A dipstick ELISA for rapid detection of human blood meals in mosquitoes. J Am Mosq Control Assoc. 1991 Mar;7(1):16–23. [PubMed] [Google Scholar]
  14. Sumathy S., Thyagarajan S. P., Latif R., Madanagopalan N., Raguram K., Rajasambandam P., Gowans E. A dipstick immunobinding enzyme-linked immunosorbent assay for serodiagnosis of hepatitis B and delta virus infections. J Virol Methods. 1992 Jul;38(1):145–152. doi: 10.1016/0166-0934(92)90177-f. [DOI] [PubMed] [Google Scholar]
  15. Weiss A. J., McElhinney S. A., Blankstein L. A. The catalysis of protein and nucleic acid coupling to an affinity membrane substrate. Biotechniques. 1989 Oct;7(9):1012–1016. [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES