Skip to main content
PMC home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1984 May;47(5):1047–1053. doi: 10.1128/aem.47.5.1047-1053.1984

Rapid enumeration of Staphylococcus aureus in foods by direct demonstration of enterotoxigenic colonies on membrane filters by enzyme immunoassay.

P I Peterkin, A N Sharpe
PMCID: PMC240052  PMID: 6742823

Abstract

Based on enzyme-linked immunosorbent assay, a convenient method has been devised for the direct demonstration of enterotoxin B production by Staphylococcus aureus colonies grown for 24 h on membrane filters. The problem of false-positive reactions due to binding of immunoglobulin G to protein A was turned to advantage by conjugating horseradish peroxidase directly to protein A, which then mediated the labeling of the antitoxin. The test requires 3 h to complete and yields a purple stain at the site of enterotoxin B-producing colonies, thus allowing direct enumeration of confirmed S. aureus in foods within 27 h. The method should be applicable to other enterotoxins of S. aureus.

Full text

PDF

Images in this article

1050Image
on p.1050
1051Image
on p.1051
1052Image
on p.1052

Selected References

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

  1. Broome S., Gilbert W. Immunological screening method to detect specific translation products. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2746–2749. doi: 10.1073/pnas.75.6.2746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Calderon R. L., Levin M. A. Quantitative method for enumeration of enterotoxigenic Escherichia coli. J Clin Microbiol. 1981 Jan;13(1):130–134. doi: 10.1128/jcm.13.1.130-134.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dubois-Dalcq M., McFarland H., McFarlin D. Protein A-peroxidase: a valluable tool for the localization of antigens. J Histochem Cytochem. 1977 Nov;25(11):1201–1206. doi: 10.1177/25.11.199666. [DOI] [PubMed] [Google Scholar]
  4. Edwards E. A., Larson G. L. New method of grouping beta-hemolytic streptococci directly on sheep blood agar plates by coagglutination of specifically sensitized protein A-containing staphylococci. Appl Microbiol. 1974 Dec;28(6):972–976. doi: 10.1128/am.28.6.972-976.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Entis P. Enumeration of coliforms in nonfat dry milk and canned custard by hydrophobic grid membrane filter method: collaborative study. J Assoc Off Anal Chem. 1983 Jul;66(4):897–904. [PubMed] [Google Scholar]
  6. Erlich H. A., Cohen S. N., McDevitt H. O. A sensitive radioimmunoassay for detecting products translated from cloned DNA fragments. Cell. 1978 Apr;13(4):681–689. doi: 10.1016/0092-8674(78)90218-0. [DOI] [PubMed] [Google Scholar]
  7. Jarvis A. W., Lawrence R. C. Production of high titers of enterotoxins for the routine testing of staphylococci. Appl Microbiol. 1970 Apr;19(4):698–699. doi: 10.1128/am.19.4.698-699.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Lachica R. V., Genigeorgis C., Hoeprich P. D. Metachromatic agar-diffusion methods for detecting staphylococcal nuclease activity. Appl Microbiol. 1971 Apr;21(4):585–587. doi: 10.1128/am.21.4.585-587.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. McLean R. A., Lilly H. D., Alford J. A. Effects of meat-curing salts and temperature on production of staphylococcal enterotoxin B. J Bacteriol. 1968 Apr;95(4):1207–1211. doi: 10.1128/jb.95.4.1207-1211.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Park C. E., de Melo Serrano A., Landgraf M., Huang J. C., Stankiewicz Z., Rayman M. K. A survey of microorganisms for thermonuclease production. Can J Microbiol. 1980 Apr;26(4):532–538. doi: 10.1139/m80-089. [DOI] [PubMed] [Google Scholar]
  11. Rayman M. K., Devoyod J. J., Purvis U., Kusch D., Lanier J., Gilbert R. J., Till D. G., Jarvis G. A. ICMSF methods studies. X. An international comparative study of four media for the enumeration of Staphylococcus aureus in foods. Can J Microbiol. 1978 Mar;24(3):274–281. doi: 10.1139/m78-047. [DOI] [PubMed] [Google Scholar]
  12. Sharpe A. N., Michaud G. L. Hydrophobic grid-membrane filters: new approach to microbiological enumeration. Appl Microbiol. 1974 Aug;28(2):223–225. doi: 10.1128/am.28.2.223-225.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sharpe A. N., Peterkin P. I., Dudas I. Membrane filtration of food suspensions. Appl Environ Microbiol. 1979 Jan;37(1):21–35. doi: 10.1128/aem.37.1.21-35.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sharpe A. N., Peterkin P. I., Malik N. Improved detection of coliforms and Escherichia coli in foods by a membrane filter method. Appl Environ Microbiol. 1979 Sep;38(3):431–435. doi: 10.1128/aem.38.3.431-435.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Sharpe A. N., Peterkin P. I., Rayman M. K. Detection of Escherichia coli in foods: indole staining methods for cellulosic and polysulfone membrane filters. Appl Environ Microbiol. 1981 Jun;41(6):1310–1315. doi: 10.1128/aem.41.6.1310-1315.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES