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Journal of Clinical Microbiology, June 2007, p. 1867-1873, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02100-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

StaphPlex System for Rapid and Simultaneous Identification of Antibiotic Resistance Determinants and Panton-Valentine Leukocidin Detection of Staphylococci from Positive Blood Cultures{triangledown}

Yi-Wei Tang,1,2* Abdullah Kilic,1,{dagger} Qunying Yang,3 Sigrid K. McAllister,4 Haijing Li,1 Rebecca S. Miller,3 Melinda McCormac,2 Karen D. Tracy,3 Charles W. Stratton,1,2 Jian Han,3 and Brandi Limbago4

Departments of Medicine,1 Pathology, Vanderbilt University Medical Center, Nashville, Tennessee 37232,2 Genaco Biomedical Products, Inc., Huntsville, Alabama 35805,3 Division of Healthcare Quality Promotion, Centers for Disease Control and Prevention, Atlanta, Georgia 303334

Received 12 October 2006/ Returned for modification 4 December 2006/ Accepted 9 April 2007

Phenotypic methods take several days for identification and antimicrobial susceptibility testing of staphylococcal isolates after gram-positive cocci in clusters (GPCC) are observed in positive blood cultures. We developed and validated a StaphPlex system that amplifies and detects 18 gene targets simultaneously in 1 reaction for species-level identification of staphylococci, detection of genes encoding Panton-Valentine leukocidin (PVL), and antimicrobial resistance determinants of staphylococci. The StaphPlex system was compared to phenotypic methods for organism identification and antimicrobial resistance detection for positive blood culture specimens in which GPCC were observed. Among a total of 360 GPCC specimens, 273 (75.8%), 37 (10.3%), 37 (10.3%), 1 (0.3%), 3 (0.8%), and 9 (2.5%) were identified by StaphPlex as coagulase-negative Staphylococcus (CoNS), methicillin-resistant Staphylococcus aureus (MRSA), methicillin-susceptible S. aureus (MSSA), or mixed infections of CoNS and MRSA, CoNS and MSSA, or nonstaphylococci, respectively, with an overall accuracy of 91.7%. The 277 CoNS-containing specimens were further identified to the species level as containing 203 (73.3%) Staphylococcus epidermidis isolates, 10 (3.6%) Staphylococcus haemolyticus isolates, 27 (9.7%) Staphylococcus hominis isolates, 1 (0.4%) Staphylococcus lugdunensis isolate, and 36 (13.0%) other CoNS isolates, with an overall accuracy of 80.1% compared to an API STAPH test and CDC reference identification. Numerous very major errors were noticed when detection of aacA, ermA, ermC, tetM, and tetK was used to predict in vitro antimicrobial resistance, but relatively few major errors were observed when the absence of these genes was used to predict susceptibility. The StaphPlex system demonstrated 100% sensitivity and specificity, ranging from 95.5% to 100.0% when used for staphylococcal cassette chromosome mec typing and PVL detection. StaphPlex provides simultaneous staphylococcal identification and detection of PVL and antimicrobial resistance determinants within 5 h, significantly shortening the time needed for phenotypic identification and antimicrobial susceptibility testing.

* Corresponding author. Mailing address: Molecular Infectious Disease Laboratory, Vanderbilt University Hospital, 4605 TVC, Nashville, TN 37232-5310. Phone: (615) 322-2035. Fax: (615) 343-8420. E-mail: yiwei.tang{at}vanderbilt.edu

{triangledown} Published ahead of print on 19 April 2007.

{dagger} Present address: Gulhane Military Medical Academy, Ankara 06018, Turkey.

Journal of Clinical Microbiology, June 2007, p. 1867-1873, Vol. 45, No. 6
0095-1137/07/$08.00+0     doi:10.1128/JCM.02100-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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