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Journal of Clinical Microbiology, September 2000, p. 3280-3284, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Development of a Rapid PCR Assay Specific for Staphylococcus saprophyticus and Application to Direct Detection from Urine Samples

Francis Martineau,1,2 François J. Picard,1 Christian Ménard,1 Paul H. Roy,1,3 Marc Ouellette,1,2 and Michel G. Bergeron1,2,*

Centre de Recherche en Infectiologie, Centre Hospitalier Universitaire de Québec (Pavillon Centre Hospitalier de l'Université Laval),1 and Division de Microbiologie, Faculté de Médecine, Université Laval,2 Ste-Foy, Québec, Canada G1V 4G2, and Département de Biochimie, Université Laval, Ste-Foy, Québec, Canada G1K 7P43

Received 29 March 2000/Returned for modification 26 May 2000/Accepted 17 June 2000

Staphylococcus saprophyticus is one of the most frequently encountered microorganisms associated with acute urinary tract infections (UTIs) in young, sexually active female outpatients. Conventional identification methods based on biochemical characteristics can efficiently identify S. saprophyticus, but the rapidities of these methods need to be improved. Rapid and direct identification of this bacterium from urine samples would be useful to improve time required for the diagnosis of S. saprophyticus infections in the clinical microbiology laboratory. We have developed a PCR-based assay for the specific detection of S. saprophyticus. An arbitrarily primed PCR amplification product of 380 bp specific for S. saprophyticus was sequenced and used to design a set of S. saprophyticus-specific PCR amplification primers. The PCR assay was specific for S. saprophyticus when tested with DNA from 49 gram-positive and 31 gram-negative bacterial species. This assay was also able to amplify efficiently DNA from all 60 strains of S. saprophyticus from various origins tested. This assay was adapted for direct detection from urine samples. The sensitivity levels achieved with urine samples was 19 CFU with 30 cycles of amplification and 0.5 CFU with 40 cycles of amplification. This PCR assay for the specific detection of S. saprophyticus is simple and rapid (approximately 90 min, including the time for urine specimen preparation).

* Corresponding author. Mailing address: Centre de Recherche en Infectiologie, CHUQ (Pavillon CHUL), 2705 Boul. Laurier, Ste-Foy, Québec, Canada G1V 4G2. Phone: (418) 654-2705. Fax: (418) 654-2715. E-mail: Michel.G.Bergeron{at}crchul.ulaval.ca.

Journal of Clinical Microbiology, September 2000, p. 3280-3284, Vol. 38, No. 9
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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