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Journal of Clinical Microbiology, November 2009, p. 3472-3477, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.00342-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Comparison of Real-Time PCR and Conventional Biochemical Methods for Identification of Staphylococcus lugdunensis{triangledown}

Benjamin A. Pinsky,1* Divinia Samson,2 Laleh Ghafghaichi,2 Ellen J. Baron,1,2 and Niaz Banaei1,2

Department of Pathology, Stanford University School of Medicine, Stanford, California 94305,1 Clinical Microbiology Laboratory, Stanford Hospital and Clinics, Palo Alto, California 943042

Received 15 February 2009/ Returned for modification 13 July 2009/ Accepted 1 September 2009

Staphylococcus lugdunensis is an aggressive, virulent member of the coagulase-negative staphylococci (CoNS) that is responsible for severe, rapidly progressive skin and soft tissue infections and native valve endocarditis. To facilitate prompt identification and appropriate therapy, we describe here a rapid and robust multiplex real-time PCR assay that is able to definitively distinguish S. lugdunensis from other staphylococci. Using melting curve analysis, the assay also identifies Staphylococcus aureus and CoNS other than S. lugdunensis and determines MecA-dependent resistance to methicillin (meticillin). When applied to a panel of well-characterized staphylococcal reference strains, as well as 165 clinical isolates previously identified by conventional methods, the assay was both sensitive and specific for S. lugdunensis, correctly identifying the reference strain and all 47 S. lugdunensis isolates without inappropriate amplification of other staphylococci. Furthermore, rapid biochemical identification using the WEE-TAB system to detect ornithine decarboxylase activity was found to be unsuitable as an alternative to PCR identification, displaying just 31% sensitivity and 77% specificity when tested on a subset (90 isolates) of the clinical strains. We therefore propose that this simple, accurate PCR approach will allow for the routine and timely identification of S. lugdunensis in the clinical microbiology laboratory.

* Corresponding author. Mailing address: Stanford University School of Medicine, Department of Pathology, 300 Pasteur Drive, Lane 235, Stanford, CA 94305-5324. Phone: (650) 723-5252. Fax: (650) 725-6902. E-mail: bpinsky{at}stanford.edu

{triangledown} Published ahead of print on 9 September 2009.

Journal of Clinical Microbiology, November 2009, p. 3472-3477, Vol. 47, No. 11
0095-1137/09/$08.00+0     doi:10.1128/JCM.00342-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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