Development of a PCR test to differentiate between Staphylococcus aureus and Staphylococcus intermedius

• Published in: Journal of food protection Vol. 67; no. 10; p. 2302
• Main Authors: Baron, Florence, Cochet, Marie-Françoise, Pellerin, Jean-louis, Ben Zakour, Nouri, Lebon, Anne, Navarro, Anne, Proudy, Isabelle, Le Loir, Yves, Gautier, Michel
• Format: Journal Article
• Language: English
• Published: United States 01-10-2004
• Subjects: Animals; DNA Primers; DNA, Bacterial - isolation & purification; Enterotoxins - biosynthesis; Food Microbiology; Gene Amplification; Genes, Bacterial; Polymerase Chain Reaction - methods; RNA, Ribosomal, 16S - analysis; Sensitivity and Specificity; Species Specificity; Staphylococcus - classification; Staphylococcus - isolation & purification; Staphylococcus - metabolism; Staphylococcus aureus - classification; Staphylococcus aureus - isolation & purification; Staphylococcus aureus - metabolism
• ISSN: 0362-028X
• DOI: 10.4315/0362-028X-67.10.2302

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.