Development of a PCR test to differentiate between Staphylococcus aureus and Staphylococcus intermedius.
The presence of Staphylococcus intermedius in food remains unclear because routine laboratory analysis does not discriminate between S. intermedius and Staphylococcus aureus, a major cause of food poisoning. Both species share many phenotypic characteristics, including coagulase and thermonuclease production. In both species, some strains can produce enterotoxin and therefore can be the cause of food poisoning outbreaks. Although the ID32 Staph System (bioMérieux, SA, Marcy l'Etoile, France), based on a miniaturized phenotypic characterization, gives satisfactory results for discriminating between these two species, some rapid molecular PCR-based methods have been developed to identify S. aureus specifically, but they do not identify S. intermedius. Here, we developed a rapid, accurate, and discriminative multiplex PCR method that targets species-specific sequences in the nuc gene, which encodes thermonuclease in the two species. The test includes an internal positive control that targets a highly conserved region of 16S ribosomal RNA gene (rDNA). A total of 116 strains were used to validate our test. The test gave no signal on the following Staphylococcus species: S. epidermidis, S. chromogenes, S. hyicus, S. warneri, S. xylosus, S. lentus, and S. sciuri. It allowed a 100% successful discrimination between S. aureus (44 strains tested) and S. intermedius (57 strains) isolated from different origins.
Baron
Florence
F
Laboratoire de Microbiologie, UMR 1055, Ecole Nationale Supérieure Agronomique, Institut National de la Recherche Agronomique, CS84215, 35042 Rennes, France. baron@agrorennes.educagri.fr
Cochet
Marie-Françoise
MF
Pellerin
Jean-louis
JL
Ben Zakour
Nouri
N
Lebon
Anne
A
Navarro
Anne
A
Proudy
Isabelle
I
Le Loir
Yves
Y
Gautier
Michel
M
D000818:Animals; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D004768:Enterotoxins / Q000096:biosynthesis; D005516:Food Microbiology; D005784:Gene Amplification; D005798:Genes, Bacterial; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013210:Staphylococcus / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism; D013211:Staphylococcus aureus / Q000145:classification / Q000302:isolation & purification* / Q000378:metabolism
D017931:DNA Primers; D004269:DNA, Bacterial; D004768:Enterotoxins; D012336:RNA, Ribosomal, 16S