Detection and genotyping by real-time PCR of the staphylococcal enterotoxin genes sea to sej

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• 作者：Letertre ; Capucine ; Perelle ; Sylvie ; Dilasser ; Franç ; oise ; Fach ; Patrick
• 关键词：Staphylococcus aureus ; Real-time PCR ; Enterotoxin
• 刊名：Molecular and Cellular Probes
• 出版年：2003
• 出版时间：August, 2003
• 年：2003
• 卷：17
• 期：4
• 页码：139-147
• 全文大小：202 K

文摘

This paper describes real-time fluorescence PCR assays for detecting and toxinotyping nine enterotoxin genes from Staphylococcus aureus. A universal set of primers allowed sea, seb, sec, sed, see, seg, seh, sei, sej enterotoxin genes from S. aureus to be detected in a single real-time PCR assay with the LightCycler (LC) instrument. Using the universal forward primer and a type-specific reverse primer, real-time PCR assays allowed the S. aureus enterotoxin genes to be specifically genotyped. A collection of S. aureus isolates (n=83) was detected and further characterised for sea, seb, sec, sed, see, seg, seh, sei, sej, using real-time PCR assays, and data were compared with those obtained by conventional block cycler PCR. Isolates were also tested for their ability to produce staphylococcal enterotoxins A, B, C and D by a commercial reversed passive latex agglutination (RPLA) test. Real-time PCR assays developed on the LightCycler system (LC-PCR) are a powerful tool for rapid detection and toxinotyping of the enterotoxin genes sea to sej from S. aureus. The work offers a very quick, reliable and specific alternative to conventional block cycler PCR assays to identify the enterotoxin profile of toxigenic S. aureus.