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Journal of Clinical Microbiology, April 2000, p. 1385-1389, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Rapid Enzyme Immunoassay for Determination of Toxigenicity among Clinical Isolates of Corynebacteria

Kathryn H. Engler* and Androulla Efstratiou

Respiratory and Systemic Infection Laboratory, Central Public Health Laboratory, London NW9 5HT, United Kingdom

Received 11 August 1999/Returned for modification 3 November 1999/Accepted 17 January 2000

A rapid enzyme immunoassay (EIA) was developed for the phenotypic detection of diphtheria toxin among clinical isolates of corynebacteria. The assay uses equine polyclonal antitoxin as the capture antibody and an alkaline phosphatase-labeled monoclonal antibody, specific for fragment A of the toxin molecule, as the detecting antibody. The assay is rapid, sensitive, and specific: a final result is available within 3 h of colony selection, and the limits of detection are 0.1 ng of pure diphtheria toxin/ml. Toxigenicity could be detected with isolates grown on a diverse range of culture media, including selective agars. Toxin detection using the EIA was compared to that with the Elek test and PCR detection of fragment A of the diphtheria toxin (tox) gene, using 245 isolates of corynebacteria. The results for the EIA were in complete concordance with those of the Elek test: 87 toxigenic and 158 nontoxigenic isolates. Ten of the phenotypically nontoxigenic strains were found to contain fragment A of the tox gene but did not express the toxin protein. These isolates were found to be nontoxigenic in the Vero cell tissue culture cytotoxicity assay and were therefore nontoxigenic for diagnostic purposes. The EIA is a simple rapid phenotypic test which provides a definitive result on toxigenicity within one working day.

* Corresponding author. Mailing address: WHO Collaborating Centre for Diphtheria and Streptococcal Infections, Respiratory and Systemic Infection Laboratory, Central Public Health Laboratory, 61 Colindale Ave., London NW9 5HT, United Kingdom. Phone: 44 181 200 4400. Fax: 44 181 205 6528. E-mail: kengler{at}phls.nhs.uk.

Journal of Clinical Microbiology, April 2000, p. 1385-1389, Vol. 38, No. 4
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

  • Bachran, C., Sutherland, M., Bachran, D., Fuchs, H. (2007). Quantification of Diphtheria Toxin Mediated ADP-Ribosylation in a Solid-Phase Assay. Clin. Chem. 53: 1676-1683 [Abstract] [Full Text]  
  • Engler, K. H., Efstratiou, A., Norn, D., Kozlov, R. S., Selga, I., Glushkevich, T. G., Tam, M., Melnikov, V. G., Mazurova, I. K., Kim, V. E., Tseneva, G. Y., Titov, L. P., George, R. C. (2002). Immunochromatographic Strip Test for Rapid Detection of Diphtheria Toxin: Description and Multicenter Evaluation in Areas of Low and High Prevalence of Diphtheria. J. Clin. Microbiol. 40: 80-83 [Abstract] [Full Text]  
  • ENGLER, K. H., KOZLOV, R. S., COPPING, S. J., DIPHTHERIA, M. O. T. E. L. W. G. O., EFSTRATIOU, A. (2001). International external quality assessment scheme for the laboratory diagnosis of diphtheria. J Med Microbiol 50: 1006-1012 [Abstract] [Full Text]  


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