Detection and genotyping by real-time PCR of the staphylococcal enterotoxin genes sea to sej.
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.
Letertre
Capucine
C
Unité: Atelier de Biotechnologie, Laboratoire d'Etudes et de Recherches sur l'Hygiène et la Qualité des Aliments, Agence Française de Sécurité Sanitaire des Aliments (AFSSA), 1-5 rue de Belfort, 94700, Maisons-Alfort, France.
Perelle
Sylvie
S
Dilasser
Françoise
F
Fach
Patrick
P
D001426:Bacterial Proteins / Q000235:genetics*; D001431:Bacteriological Techniques; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D004768:Enterotoxins / Q000145:classification / Q000235:genetics*; D005516:Food Microbiology; D005798:Genes, Bacterial; D005838:Genotype; D006801:Humans; D007841:Latex Fixation Tests / Q000379:methods; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D004768:Enterotoxins