Identification and measurement of staphylococcal enterotoxin-like protein I (SEll) secretion from Staphylococcus aureus clinical isolate.

AIMS: Staphylococcus aureus (Staph. aureus) produces a wide variety of staphylococcal enterotoxins (SEs) and staphylococcal enterotoxin-like (SEl) proteins, which are the most causative agents of staphylococcal food poisoning. In contrast to classical SEs (SEA to SEE), the relationship between the novel SEs/SEls (SEG to SElX) and staphylococcal food poisoning is not elucidated. This study is aimed to establish a system to detect staphylococcal enterotoxin-like protein I (SElI) for analysis of staphylococcal food poisoning.

METHODS AND

RESULTS

SElI was characterized in a Staph. aureus clinical isolate associated with food poisoning; there was an amino acid substitution Thr145Ala compared to previously identified SEI from Staph. aureus 04-02981. Subsequently, SElI was expressed, purified, and the poly- and monoclonal antibodies against it were prepared. Using these antibodies, a highly sensitive sandwich enzyme-linked immunosorbent assay (ELISA) that specifically detected and measured SElI secretion from the Staph. aureus clinical isolate in LB medium, milk and bloodstream was developed.

CONCLUSIONS

The ELISA system has been successfully applied for analysing SElI secretion in vivo and in vitro.

SIGNIFICANCE AND IMPACT OF THE STUDY: The highly sensitive ELISA should make it attractive for quantifying SElI in food hygiene supervision and clinical diagnosis in near future.