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Simultaneous analysis of multiple staphylococcal enterotoxin genes by an oligonucleotide microarray assay
Nikolay Sergeev1, Dmitriy Volokhov, Vladimir Chizhikov
1Center for Food Safety and Applied Nutrition, Food and Drug Administration, College Park, Maryland, USA.
Journal of Clinical Microbiology
|May 8, 2004
Summary
A new microarray assay enables simultaneous detection of multiple staphylococcal enterotoxin (SE) genes in Staphylococcus aureus. This rapid method improves identification of foodborne illness-causing bacteria and potential bioweapons.
Area of Science:
- Microbiology
- Molecular Biology
- Food Safety
Background:
- Staphylococcal enterotoxins (SEs) cause foodborne gastroenteritis and are potential bioweapons.
- Current typing methods (serological, PCR) for SEs are time-consuming and may be inaccurate.
- Many Staphylococcus aureus strains harbor multiple enterotoxin genes.
Purpose of the Study:
- To develop a rapid, simultaneous assay for detecting and genetically typing multiple enterotoxin (ent) genes.
- To overcome limitations of existing serological and PCR-based typing methods for SEs.
Main Methods:
- A one-tube microarray-based assay using PCR amplification with degenerate primers.
- Hybridization of PCR products with specific oligonucleotide probes for ent gene identification.
- Validation using gene-specific PCR, gel electrophoresis, microarray hybridization, and sequencing.
Main Results:
- The microarray assay successfully detected and identified multiple ent genes simultaneously in S. aureus strains.
- Identified previously undetected ent genes in some isolates.
- The assay can identify up to nine different ent genes in a single S. aureus strain.
Conclusions:
- The oligonucleotide microarray assay is effective for analyzing multitoxigenic S. aureus strains.
- This method offers a valuable tool for microbial pathogen analysis, particularly for food safety and biodefense applications.