1887

Abstract

This study evaluated fluorescence hybridization (FISH) for rapid identification of and coagulase-negative staphylococci (CoNS) directly from blood cultures. Initially, 360 blood cultures containing Gram-positive cocci were investigated by a previously described microwave-FISH procedure: 44/49 (89.8 %) and 298/299 (99.7 %) CoNS were correctly identified. Because FISH proved useful and reliable but handling was found to be inconvenient, the method was modified by employing a recently developed slide chamber. This reduced the time required from 60 to 30 min. The simplified execution allowed integration of the method into the workflow of a routine laboratory without difficulty. The modified method proved to be highly reliable, identifying 37/37 (100 %) and 169/172 (98.2 %) CoNS directly from blood cultures.

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2010-01-01
2021-10-20
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References

  1. Amann R. I., Binder B. J., Olson R. J., Chisholm S. W., Devereux R., Stahl D. A. 1990; Combination of 16S rRNA-targeted oligonucleotide probes with flow cytometry for analyzing mixed microbial populations. Appl Environ Microbiol 56:1919–1925
    [Google Scholar]
  2. Barenfanger J., Drake C., Kacich G. 1999; Clinical and financial benefits of rapid bacterial identification and antimicrobial susceptibility testing. J Clin Microbiol 37:1415–1418
    [Google Scholar]
  3. Chapin K., Musgnug M. 2003; Evaluation of three rapid methods for the direct identification of Staphylococcus aureus from positive blood cultures. J Clin Microbiol 41:4324–4327 [CrossRef]
    [Google Scholar]
  4. Forrest G. N., Mehta S., Weekes E., Lincalis D. P., Johnson J. K., Venezia R. A. 2006; Impact of rapid in situ hybridization testing on coagulase-negative staphylococci positive blood cultures. J Antimicrob Chemother 58:154–158 [CrossRef]
    [Google Scholar]
  5. Gescher D. M., Kovacevic D., Schmiedel D., Siemoneit S., Mallmann C., Halle E., Göbel U. B., Moter A. 2008; Fluorescence in situ hybridisation (FISH) accelerates identification of Gram-positive cocci in positive blood cultures. Int J Antimicrob Agents 32 (Suppl. 1):S51–S59 [CrossRef]
    [Google Scholar]
  6. González V., Padilla E., Gimenez M., Vilaplana C., Pérez A., Fernández G., Quesada M. D., Pallarés M. A., Ausina V. 2004; Rapid diagnosis of Staphylococcus aureus bacteremia using S. aureus PNA FISH. Eur J Clin Microbiol Infect Dis 23:396–398 [CrossRef]
    [Google Scholar]
  7. Hartmann H., Stender H., Schafer A., Autenrieth I. B., Kempf V. A. 2005; Rapid identification of Staphylococcus aureus in blood cultures by a combination of fluorescence in situ hybridization using peptide nucleic acid probes and flow cytometry. J Clin Microbiol 43:4855–4857 [CrossRef]
    [Google Scholar]
  8. Hensley D. M., Tapia R., Encina Y. 2009; An evaluation of the AdvanDx Staphylococcus aureus /CNS PNA FISH assay. Clin Lab Sci 22:30–33
    [Google Scholar]
  9. Hiraishi A. 1992; Direct automated sequencing of 16S rDNA amplified by polymerase chain reaction from bacterial cultures without DNA purification. Lett Appl Microbiol 15:210–213 [CrossRef]
    [Google Scholar]
  10. Jansen G. J., Mooibroek M., Idema J., Harmsen H. J., Welling G. W., Degener J. E. 2000; Rapid identification of bacteria in blood cultures by using fluorescently labeled oligonucleotide probes. J Clin Microbiol 38:814–817
    [Google Scholar]
  11. Kempf V. A., Trebesius K., Autenrieth I. B. 2000; Fluorescent in situ hybridization allows rapid identification of microorganisms in blood cultures. J Clin Microbiol 38:830–838
    [Google Scholar]
  12. Ly T., Gulia J., Pyrgos V., Waga M., Shoham S. 2008; Impact upon clinical outcomes of translation of PNA FISH-generated laboratory data from the clinical microbiology bench to bedside in real time. Ther Clin Risk Manag 4:637–640
    [Google Scholar]
  13. Mehta M. S., Paule S. M., Thomson R. B., Kaul K. L., Peterson L. R. 2009; Identification of Staphylococcus species directly from positive blood culture broth by use of molecular and conventional methods. J Clin Microbiol 47:1082–1086 [CrossRef]
    [Google Scholar]
  14. Moter A., Göbel U. B. 2000; Fluorescence in situ hybridization (FISH) for direct visualization of microorganisms. J Microbiol Methods 41:85–112 [CrossRef]
    [Google Scholar]
  15. Oliveira K., Procop G. W., Wilson D., Coull J., Stender H. 2002; Rapid identification of Staphylococcus aureus directly from blood cultures by fluorescence in situ hybridization with peptide nucleic acid probes. J Clin Microbiol 40:247–251 [CrossRef]
    [Google Scholar]
  16. Oliveira K., Brecher S. M., Durbin A., Shapiro D. S., Schwartz D. R., De Girolami P. C., Dakos J., Procop G. W., Wilson D. other authors 2003; Direct identification of Staphylococcus aureus from positive blood culture bottles. J Clin Microbiol 41:889–891 [CrossRef]
    [Google Scholar]
  17. Peters R. P., van Agtmael M. A., Simoons-Smit A. M., Danner S. A., Vandenbroucke-Grauls C. M., Savelkoul P. H. 2006; Rapid identification of pathogens in blood cultures with a modified fluorescence in situ hybridization assay. J Clin Microbiol 44:4186–4188 [CrossRef]
    [Google Scholar]
  18. Ruimy R., Dos-Santos M., Raskine L., Bert F., Masson R., Elbaz S., Bonnal C., Lucet J. C., Lefort A. other authors 2008; 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. J Clin Microbiol 46:2045–2051 [CrossRef]
    [Google Scholar]
  19. Speers D. J., Olma T. R., Gilbert G. L. 1998; Evaluation of four methods for rapid identification of Staphylococcus aureus from blood cultures. J Clin Microbiol 36:1032–1034
    [Google Scholar]
  20. Tavares A., Inacio J., Melo-Cristino J., Couto I. 2008; Use of fluorescence in situ hybridization for rapid identification of staphylococci in blood culture samples collected in a Portuguese hospital. J Clin Microbiol 46:3097–3100 [CrossRef]
    [Google Scholar]
  21. Weinstein M. P. 2003; Blood culture contamination: persisting problems and partial progress. J Clin Microbiol 41:2275–2278 [CrossRef]
    [Google Scholar]
  22. Wellinghausen N., Bartel M., Essig A., Poppert S. 2007; Rapid identification of clinically relevant Enterococcus species by fluorescence in situ hybridization. J Clin Microbiol 45:3424–3426 [CrossRef]
    [Google Scholar]
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