1887

Microbiology Society logo

Volume 140, Issue 12

Detection of by ELISA utilizing antibodies produced against its exoantigens Free

Abstract

An indirect competitive ELISA was developed using rabbit antiserum against the exoantigens (ExAgs) of , and cross-reactivities were determined with 15 ExAgs from other species and genera of fungi and with the water-soluble extracts from four grains. The assay had good sensitivity to ExAgs (1 μg per ml serum), with generally low or no cross-reactivity with grain extracts and the other fungi, including five species of , four species of , three species of , two species of and one species of Immunoblotting analysis of the ExAgs from yielded at least 20 distinct antigenic bands, with about 3 or 4 being dark, 10-12 light and 5-8 faint in colour. Most of the bands had molecular masses of about 70-90 kDa. In contrast to , the ExAgs from the other fungi either did not react in the immunoblotting assay with the antiserum, or reacted only weakly with it. One antigenic band from , however, reacted moderately strongly with the antiserum. The ELISA and immunoblotting analysis were used to detect in naturally contaminated wheat samples, and in wheat samples spiked with a second common storage fungus, The results demonstrated that the antiserum was able to detect in a background of other fungal species, and that these did not produce false positives. There was also a positive association between the concentration of ExAgs, as measured by ELISA, and the amount of mould as determined by the number of colony-forming units. Immunoblotting qualitatively confirmed the ELISA results obtained with fungi when cultured on liquid or on solid (wheat) media. The data suggest that the immunoassays developed for are useful for the detection and identification of on the basis of its ExAgs, with the advantages of being more efficient, simple and reliable than conventional techniques.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): ELISA , exoantigens , immunoblotting and Penicillium aurantiogriseum
© Society for General Microbiology, 1994
Loading

Article metrics loading...

/content/journal/micro/10.1099/13500872-140-12-3267
1994-12-01
2024-04-25
Loading full text...

Full text loading...

/deliver/fulltext/micro/140/12/mic-140-12-3267.html?itemId=/content/journal/micro/10.1099/13500872-140-12-3267&mimeType=html&fmt=ahah

References

  1. Banks J.N., Cox S.J., Northway B.J. Polyclonal and monoclonal antibodies to field and storage fungi. Int Biodetenor & Biodegrad 1993; 32:137–144
     [Google Scholar]
  2. Blake M.S., Johnstone K.H., Russell-Jones G.J., Gotschlich E.C. A rapid, sensitive method for detection of alkaline phosphatase-conjugated anti-antibody on Western blots. Anal Biochem 1984; 136:175–179
     [Google Scholar]
  3. Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976; 72:248–254
     [Google Scholar]
  4. CAST Mjcotoxins: Economic and Health Risks 1989 Ames, Iowa: Council for Agricultural Science and Technology.;
     [Google Scholar]
  5. Clarke J.R., Marquardt R.R., Oosterveld A., Frohlich A.A. Development of a quantitative and sensitive enzyme-linked immunoassay for ochratoxin A using antibodies from the yolk of the laying hen. J Agric Food Chem 1993; 41:1784–1789
     [Google Scholar]
  6. Dewey F.M., MacDonald M.M., Phillips S.I., Priestley R.A. Development of monoclonal-antibody-ELISA and-DIPSTICK immunoassays for Penicillium islandicum in rice grains. J Gen Microbzoll 1990; 36:753–760
     [Google Scholar]
  7. Dubois M., Gilles K.A., Hamilton J.K., Rebers P.A., Smith F. Colorimetric method for determination of sugars and related substances. Anal Chem 1956; 28:350–356
     [Google Scholar]
  8. Frisvad J.C. Fungal species and their specific production of mycotoxins. In Introduction to Food-borne Fungi 1988 Edited by Samson R.A., Van Reenen-Hoekstra E. Baarn: Centralbureau Voor Schimmelcultures; pp 239–249
     [Google Scholar]
  9. Frisvad J.C. The connection between the penicillia and aspergilli and mycotoxins with special emphasis on misidentified isolates. Arch Environ Contam Toxicol 1989; 18:452–467
     [Google Scholar]
  10. Frisvad J.C., Filtenborg O. Terverticillate Penicillia: chemotaxonomy and mycotoxin production. Mycologia 1989; 81:837–861
     [Google Scholar]
  11. Fuhrmann B., Roquebert M.F., Lebreton V., Van Hoegaerden M. Immunological differentiation between Penicillium and Aspergillus taxa. In Modern Concepts in Penicillium and Aspergillus Classification 1990 Edited by Samson R.A., Pitt J.I. New York: Plenum Press; pp 423–432
     [Google Scholar]
  12. Fuhrmann B., Lebreton V., Van Hoegaerden M., Kamphuis H.J., Strosberg A.D. A monoclonal antibody specific for conidia and mycelium wall layer of Penicillium and Aspergillus. Microbiol Immunol 1992; 36:1–12
     [Google Scholar]
  13. Harlow E., Lane D. Antibodies: a Laboratorj Manual 1988 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
     [Google Scholar]
  14. Hearn V.M., Wilson E.V., Latge J.-P. Immunological studies on Aspergillusfumigatus mycelial antigens by polyacrylamide gel electrophoresis and Western blotting techniques. J Gen Microbiol 1990; 136:1525–1535
     [Google Scholar]
  15. International Organization for Standardization (ISO) 6887E Microbiology-general guidance for the preparation of dilutions for microbiological examination 1983a
     [Google Scholar]
  16. International Organization for Standardization (ISO) TC34/SC4/WG2 Grain, seeds and derived products-Microbiological examination-Colony count technique 1983b
     [Google Scholar]
  17. Jarvis B., Williams A.P. Methods for detecting fungi in foods and beverages. In Food and Beverage Mycology 1987 Edited by Beuchat L.R. New York: Van Nostrand Reinhold; 2nd edn, pp 599–636
     [Google Scholar]
  18. Jarvis B., Seiler D.A.L., Ould A.J.L., Williams A.P. Observations on the enumerations of molds in food and feedingstuffs. J Appl Bacteriol 1983; 55:325–336
     [Google Scholar]
  19. Kaufman L., Standard P.G. Specific and rapid identification of medically important fungi by exoantigen detection. Annu Rev Microbiol 1987; 41:209–225
     [Google Scholar]
  20. Laemmli U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680–685
     [Google Scholar]
  21. Lin H.H., Lister R.M., Cousin M.A. Enzyme-linked immunosorbent assay for detection of mold in tomato puree. J Food Sci 1986; 51:180–192
     [Google Scholar]
  22. Lu P., Marquardt R.R., Frohlich A.A., Mills J. The development of a solid-phase immunoassay for the detection and identification of Aspergillus ochraceus. Food Agric Immunol 1994 (in press)
    [Google Scholar]
  23. Lund F., Frisvad J.C. Chemotaxonomy of Penicillium aurantiogriseum and related species. Mycol Res 1994 (in press)
    [Google Scholar]
  24. MacDonald M.M., Dunstan R.H., Dewey F.M. Detection of low-Air glycoproteins in surface washes of some fungal cultures by gel-filtration HPLC and by monoclonal antibodies. J Gen Microbiol 1989; l35:375–383
     [Google Scholar]
  25. Mills J.T., Wallace H.A.H. Microflora and condition of cereal seeds after a wet harvest. Can J Plant Sci 1979; 59:645–651
     [Google Scholar]
  26. Mills J.T., Frisvad J.C., Seifert K.A., Abramson D. Identification of nephrotoxic Penicillium species from cereal grains. Mycotoxin Res 1994 (in press)
    [Google Scholar]
  27. Noterrnans S., Heuvelman C.J. Immunological detection of molds in food by using the enzyme-linked immunosorbent assay (ELISA); preparation of antigens. Int J Food Microbiol 1985; 2:247–258
     [Google Scholar]
  28. Noterrnans E., Kamphuis H.J. Detection of fungi in foods by latex agglutination: a collaborative study. In Modern Methods in Food Mycology 1992 Edited by Samson R.A., Hocking A.D., Pitt J.I., King A.D. Amsterdam: Elsevier; pp 205–212
     [Google Scholar]
  29. Notermans S., Heuvelman C.J., VanEgmond H.P., Paulsch W.E., Besling J.R. Detection of mold in food by enzyme-linked immunosorbent assay. J Food Prot 1986; 49:786–791
     [Google Scholar]
  30. Pitt J.I., Hocking A.D. Fungi and Food Spoilage 1985 Sydney: Academic Press;
     [Google Scholar]
  31. Pitt J.I., Samson R.A. Approaches to Pénicillium and Aspergillus systematics. Stud Mycol 1990; 32:77–90
     [Google Scholar]
  32. Polonelli L., Castagnola M., Morace G. Identification and serotyping of Microsporum canis isolates by monoclonal antibodies. J Clin Microbiol 1986; 23:609–615
     [Google Scholar]
  33. Preston J.F., Lapis E., Gander J.E. Immunological investigations of Pénicillium. I. Serological reactivities of exocellular polysaccharides produced by six Pénicillium species. Can J Microbiol 1970; 16:687–694
     [Google Scholar]
  34. Rotter R.G., Fröhlich A.A., Marquardt R.R., Mills P.A. Estimation of fungal contamination of cereal grains as determined by measuring glucosamine concentration. Can J Anim Sei 1989; 69:235–245
     [Google Scholar]
  35. Samson R.A., Frisvad J.C. Current taxonomic concepts in Penicillium and Aspergillus. In Cereal Grains: Mycotoxins Fungi and Quality in Drying and Storage 1991 Edited by Chelkowski J. Amsterdam: Elsevier; (Dev Food Sei) 26 pp 405–439
     [Google Scholar]
  36. Standard P.G., Kaufman L., Whaley S.D. Rapid Identification of Pathogenic Mold Isolates by Immunodiffusion (CDC Lab Manual) 1985 Atlanta, GA: Centers for Disease Control;
     [Google Scholar]
  37. Tsai G.-J., Cousin M.A. Enzyme-linked immunosorbent assay for detection of molds in cheese and yogurt. J Dairy Sei 1990; 73:3366–3378
     [Google Scholar]
  38. Van Der Horst M., Samson R.A., Karman H. Comparison of two commercial kits to detect molds by latex agglutination. In Modern Methods in Food Mycology 1992 Edited by Samson R.A., Hocking A.D., Pitt J.I., King A.D. Amsterdam: Elsevier; pp 241–245
     [Google Scholar]
  39. Yu B., Niki Y., Armstrong D. Use of immunoblotting to detect Aspergillus fumigatus antigen in sera and urine of rats with experimental invasive aspergillosis. J Clin Microbiol 1990; 28:1575–1579
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/13500872-140-12-3267
Loading
Detection of Penicillium aurantiogriseumby ELISA utilizing antibodies produced against its exoantigens
Microbiology 140, 3267 (1994); https://doi.org/10.1099/13500872-140-12-3267
/content/journal/micro/10.1099/13500872-140-12-3267
/content/journal/micro/10.1099/13500872-140-12-3267
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error