2016•Journal of Applied MicrobiologyRequires access

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

Yanying Zhao, Aihua Zhu, Junni Tang, Cheng Tang, J. Chen, Jia Liu

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Abstract

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.

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What this paper is about

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.

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Available abstract

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.

Key concepts: Enterotoxin, Staphylococcus aureus, Microbiology, Micrococcaceae, Secretion, Identification (biology), Biology, Staphylococcus

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