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Journal of Clinical Microbiology, June 2008, p. 2045-2051, Vol. 46, No. 6
0095-1137/08/$08.00+0     doi:10.1128/JCM.02250-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Accuracy and Potential Usefulness of Triplex Real-Time PCR for Improving Antibiotic Treatment of Patients with Blood Cultures Showing Clustered Gram-Positive Cocci on Direct Smears{triangledown}

Raymond Ruimy,1* Marie Dos-Santos,1 Laurent Raskine,2 Frédéric Bert,3 René Masson,1 Sandrine Elbaz,1 Christine Bonnal,1 Jean-Christophe Lucet,1 Agnès Lefort,3 Bruno Fantin,3 Michel Wolff,1 Michele Hornstein,4 and Antoine Andremont1

Hospital Group Bichat-Claude Bernard, AP-HP, and EA 3964, University Denis Diderot-Paris 7 Medical School, 75870 Paris, Cedex 18, France,1 Hospital Lariboisière, AP-HP, Paris, France,2 Hospital Beaujon, AP-HP, and EA 3964, University Denis Diderot-Paris 7 Medical School, Clichy, France,3 Hospital Avicenne, AP-HP, Bobigny, France4

Received 21 November 2007/ Returned for modification 24 January 2008/ Accepted 9 April 2008

Bacterial identification and antibiotic susceptibility testing currently require 48 h when a first blood culture (BC) is positive for clustered gram-positive cocci on direct smear examination (DSE). Meanwhile, antibiotic treatment is often inadequate, reducing the chances of effective treatment or creating unnecessary selective pressure. A new real-time PCR (RT-PCR) technique that differentiates Staphylococcus aureus from coagulase-negative staphylococci (CoNS) and detects methicillin resistance in 90 min in BC bottles could help solve these problems. BC bottles from 410 patients with gram-positive cocci on DSE were processed by current methods, and patients' treatments were prospectively recorded. The RT-PCR assay was performed on aliquots of these BCs, which had been kept frozen. For the 121 patients who had true bacteremia, we established whether the faster availability of RT-PCR results could have led to the initiation of treatments different from those actually given. RT-PCR sensitivity and specificity were 100% for differentiating between S. aureus and CoNS and detecting methicillin resistance with two manufacturers' BC bottles. For 31/86 (36%) of the S. aureus-infected patients and for 8/35 (23%) of the CoNS-infected patients who either had suboptimal or nonoptimal treatment or were untreated 48 h after positivity was detected, the early availability of RT-PCR results could have allowed more effective treatment. Unnecessary glycopeptide treatments could have been avoided for 28 additional patients. The use of RT-PCR would increase treatment effectiveness in patients with staphylococcal bacteremia and reduce the selective pressure created by glycopeptides.

* Corresponding author. Mailing address: Hospital Group Bichat-Claude Bernard, Laboratoire de Bactériologie, 46, Rue Henri-Huchard, 75018 Paris, France. Phone: 33 1 40 25 85 05. Fax: 33 1 40 25 85 81. E-mail: raymond.ruimy{at}bch.ap-hop-paris.fr

{triangledown} Published ahead of print on 16 April 2008.

Journal of Clinical Microbiology, June 2008, p. 2045-2051, Vol. 46, No. 6
0095-1137/08/$08.00+0     doi:10.1128/JCM.02250-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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