Direct Identification of Staphylococcus aureus from Positive Blood Culture Bottles

Author:

Oliveira Kenneth1,Brecher Stephen M.2,Durbin Annette2,Shapiro Daniel S.3,Schwartz Donald R.3,De Girolami Paola C.4,Dakos Joanna4,Procop Gary W.5,Wilson Deborah5,Hanna Chad S.6,Haase Gerhard7,Peltroche-Llacsahuanga Heidrun7,Chapin Kimberle C.8,Musgnug Michael C.8,Levi Michael H.9,Shoemaker Cynthia9,Stender Henrik10

Affiliation:

1. Applied Biosystems, Bedford

2. Boston VA Healthcare System, West Roxbury

3. Boston Medical Center

4. Beth Israel Deaconess Medical Center, Boston, Massachusetts

5. Cleveland Clinic Foundation, Cleveland, Ohio

6. New York University School of Medicine, Bellevue Hospital, New York

7. University Hospital RWTH Aachen, Germany

8. Lahey Clinic, Burlington

9. Montefiore Medical Center, Bronx, New York

10. AdvanDx, Concord

Abstract

ABSTRACT Fluorescence in situ hybridization (FISH) using peptide nucleic acid (PNA) probes targeting Staphylococcus aureus 16S rRNA is a novel method for direct identification of S. aureus from positive blood culture bottles. The test (S. aureus PNA FISH) is performed on smears made directly from positive blood culture bottles with gram-positive cocci in clusters (GPCC) and provides results within 2.5 h. A blinded comparison of S. aureus PNA FISH with standard identification methods was performed in collaboration with eight clinical microbiology laboratories. A total of 564 routine blood culture bottles positive for GPCC recovered from both aerobic and anaerobic media from three different manufacturers (ESP, BACTEC, and BacT/Alert) were included in the study. The sensitivity and specificity of S. aureus PNA FISH were 100% (57 of 57) and 99.2% (116 of 117), respectively, with 174 GPCC-positive ESP blood culture bottles, 98.5% (67 of 68) and 98.5% (129 of 131), respectively, with 200 GPCC-positive BACTEC blood culture bottles, and 100% (74 of 74) and 99.1% (115 of 116), respectively, with 190 GPCC-positive BacT/Alert blood culture bottles. It is concluded that S. aureus PNA FISH performs well with commonly used continuously monitoring blood culture systems.

Publisher

American Society for Microbiology

Subject

Microbiology (medical)

Reference10 articles.

1. Baron E. J. 1998. Processing and interpretation of blood cultures p. 58-62. In H. D. Isenberg (ed.) Essential procedures for clinical microbiology. ASM Press Washington D.C.

2. Benito, M. J., M. M. Rodriguez, M. G. Cordoba, E. Aranda, E., and J. J. Cordoba. 2000. Rapid differentiation of Staphylococcus aureus from staphylococcal species by arbitrarily primed-polymerase chain reaction. Lett. Appl. Microbiol.31:368-373.

3. Direct identification of bacterial isolates in blood cultures by using a DNA probe

4. Egholm, M., O. Buchard, L. Christensen, C. Behrens, S. M. Freier, D. A. Driver, R. H. Berg, S. K. Kim, B. Norden, and P. E. Nielsen. 1993. PNA hybridizes to complementary oligonucleotides obeying the Watson-Crick hydrogen bonding rules. Nature365:556-568.

5. Fluorescent In Situ Hybridization Allows Rapid Identification of Microorganisms in Blood Cultures

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