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Journal of Clinical Microbiology, June 2001, p. 2050-2054, Vol. 39, No. 6
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.6.2050-2054.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Development and Evaluation of Detection Systems for Staphylococcal Exfoliative Toxin A Responsible for Scalded-Skin Syndrome

Shamez Ladhani,1,2,* Scott Robbie,1,2 Richard C. Garratt,3 Daniel S. Chapple,1,4 Christopher L. Joannou,1 and Robert W. Evans1

Metalloprotein Research Group, Division of Biomolecular Sciences, Kings College London, London SE1 9RT,1 Department of Paediatrics, Guy's Hospital, London SE1 9RT,2 and Division of Life Sciences, University of East London, London E15 4LZ,4 United Kingdom, and Instituto de Fisica, Universidade de Sao Paulo, Sao Carlos-SP, CEP 13560-970, Brazil3

Received 20 November 2000/Returned for modification 19 December 2000/Accepted 11 January 2001

Staphylococcal scalded-skin syndrome is usually diagnosed clinically by its characteristic exfoliating rash. Isolation of Staphylococcus aureus from the patient further supports the diagnosis. Several detection systems have been developed to determine whether the isolated strain produces exfoliative toxin, but none are routinely available in hospital laboratories. In a novel approach, we used computer models to predict the structure of the exfoliative toxins based on other serine proteases and to identify surface epitopes for the production of antibodies that specifically bound the exfoliative toxin A (ETA) serotype. Several rapid immunologically based diagnostic tests for ETA were developed with these antibodies and compared with existing systems. Our results showed that Western blot analysis using these antibodies was in complete correlation with PCR, which has been validated against the "gold standard" mouse model. On the other hand, the double-antibody enzyme-linked immunosorbent assay (ELISA) and Ouchterlony immunodiffusion assay gave unacceptably high false-positive results due to interference by staphylococcal protein A. This problem was successfully overcome by the development of a F(ab')2 fragment ELISA, which was rapid and reproducible and was as sensitive and specific as PCR and Western blot analysis. The F(ab')2 fragment ELISA is superior to existing diagnostic systems because it is quantitative, which may be related to the severity of the condition, and can detect amounts of exfoliative toxin in the picogram range directly from serum. This is the first detection system with the potential to confirm the diagnosis of staphylococcal scalded-skin syndrome from a routine blood test within 3 h of presentation.

* Corresponding author. Mailing address: Division of Biomolecular Sciences, 3rd Floor, New Hunts House, Guy's Hospital, London SE1 9RT, United Kingdom. Phone: 44 207 848 6482. Fax: 44 207 848 6485. E-mail: DrShamez{at}aol.com.

Journal of Clinical Microbiology, June 2001, p. 2050-2054, Vol. 39, No. 6
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.6.2050-2054.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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