1887

Abstract

Early detection of aetiological agents is pivotal for adequate therapy for bacterial infections. Although culture is still considered the mainstay for laboratory diagnosis, it often lacks sensitivity, especially in patients already treated with antibiotics. The present study investigated the potential clinical utility of the commercial real-time-PCR-based system SeptiFast (SF), originally intended for diagnosis of sepsis from blood specimens, in the aetiological diagnosis of other bacterial infections, in patients undergoing antibiotic therapy. A total of 53 non-blood specimens were analysed for microbial pathogen detection by conventional culture and with SF real-time PCR: 19 (35.8 %) synovial fluids, 9 (17.0 %) cardiac valve tissues and 25 (47.2 %) purulent exudates from various body sites. Overall, the number of specimens positive for a pathogen by SF (26/53; 49.1 %) was significantly greater ( = 0.001) than that of specimens positive by culture (10/53; 18.9 %). In particular, SF was superior to culture for pathogen detection in cardiac valve tissues and synovial fluids. The analysis of concordance showed a fair agreement between the two methods (kappa value = 0.314; 95 % confidence interval = 0.531–0.097). Even with the limitation of the low number of specimens, this study confirmed the great potential of diagnosing bacterial infections by a molecular approach, and indicates that the real-time PCR SF system can be used for specimens other than blood, from patients undergoing antibiotic treatment.

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2011-12-01
2024-04-20
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References

  1. Achermann Y., Vogt M., Leunig M., Wüst J., Trampuz A. 2010; Improved diagnosis of periprosthetic joint infection by multiplex PCR of sonication fluid from removed implants. J Clin Microbiol 48:1208–1214 [View Article][PubMed]
    [Google Scholar]
  2. Bauer M., Reinhart K. 2010; Molecular diagnostics of sepsis–where are we today?. Int J Med Microbiol 300:411–413 [View Article][PubMed]
    [Google Scholar]
  3. Costerton J. W., Post J. C., Ehrlich G. D., Hu F. Z., Kreft R., Nistico L., Kathju S., Stoodley P., Hall-Stoodley L. et al. other authors 2011; New methods for the detection of orthopedic and other biofilm infections. FEMS Immunol Med Microbiol 61:133–140 [View Article][PubMed]
    [Google Scholar]
  4. David M. Z., Daum R. S. 2010; Community-associated methicillin-resistant Staphylococcus aureus: epidemiology and clinical consequences of an emerging epidemic. Clin Microbiol Rev 23:616–687 [View Article][PubMed]
    [Google Scholar]
  5. Durack D. T., Lukes A. S., Bright D. K.Duke Endocarditis Service 1994; New criteria for diagnosis of infective endocarditis: utilization of specific echocardiographic findings. Am J Med 96:200–209 [View Article][PubMed]
    [Google Scholar]
  6. Fernández A. L., Varela E., Martínez L., Martínez A., Sierra J., González-Juanatey J. R., Regueiro B. 2010; Evaluation of a multiplex real-time PCR assay for detecting pathogens in cardiac valve tissue in patients with endocarditis. Rev Esp Cardiol 63:1205–1208 [View Article][PubMed]
    [Google Scholar]
  7. Gallo J., Kolar M., Dendis M., Loveckova Y., Sauer P., Zapletalova J., Koukalova D. 2008; Culture and PCR analysis of joint fluid in the diagnosis of prosthetic joint infection. New Microbiol 31:97–104[PubMed]
    [Google Scholar]
  8. Jorgensen J. H., Mirrett S., McDonald L. C., Murray P. R., Weinstein M. P., Fune J., Trippy C. W., Masterson M., Reller L. B. 1997; Controlled clinical laboratory comparison of BACTEC plus aerobic/F resin medium with BacT/Alert aerobic FAN medium for detection of bacteremia and fungemia. J Clin Microbiol 35:53–58[PubMed]
    [Google Scholar]
  9. Klaschik S., Lehmann L. E., Raadts A., Book M., Hoeft A., Stuber F. 2002; Real-time PCR for detection and differentiation of gram-positive and gram-negative bacteria. J Clin Microbiol 40:4304–4307 [View Article][PubMed]
    [Google Scholar]
  10. Landis J. R., Koch G. G. 1977; The measurement of observer agreement for categorical data. Biometrics 33:159–174 [View Article][PubMed]
    [Google Scholar]
  11. Lee J., Kang C. I., Lee J. H., Joung M., Moon S., Wi Y. M., Chung D. R., Ha C. W., Song J. H., Peck K. R. 2010; Risk factors for treatment failure in patients with prosthetic joint infections. J Hosp Infect 75:273–276 [View Article][PubMed]
    [Google Scholar]
  12. Lehmann L. E., Hunfeld K. P., Emrich T., Haberhausen G., Wissing H., Hoeft A., Stüber F. 2008; A multiplex real-time PCR assay for rapid detection and differentiation of 25 bacterial and fungal pathogens from whole blood samples. Med Microbiol Immunol (Berl) 197:313–324 [View Article][PubMed]
    [Google Scholar]
  13. Louie R. F., Tang Z., Albertson T. E., Cohen S., Tran N. K., Kost G. J. 2008; Multiplex polymerase chain reaction detection enhancement of bacteremia and fungemia. Crit Care Med 36:1487–1492 [View Article][PubMed]
    [Google Scholar]
  14. Mancini N., Clerici D., Diotti R., Perotti M., Ghidoli N., De Marco D., Pizzorno B., Emrich T., Burioni R. et al. other authors 2008; Molecular diagnosis of sepsis in neutropenic patients with haematological malignancies. J Med Microbiol 57:601–604 [View Article][PubMed]
    [Google Scholar]
  15. Neut D., Van der Mei H. C., Bulstra S. K., Busscher H. J. 2007; The role of small-colony variants in failure to diagnose and treat biofilm infections in orthopedics. Acta Orthop 78:299–308 [View Article][PubMed]
    [Google Scholar]
  16. Nguyen P. T., Abranches J., Phan T. N., Marquis R. E. 2002; Repressed respiration of oral streptococci grown in biofilms. Curr Microbiol 44:262–266 [View Article][PubMed]
    [Google Scholar]
  17. Peters R. P., Van Agtmael M. A., Danner S. A., Savelkoul P. H., Vandenbroucke-Grauls C. M. 2004; New developments in the diagnosis of bloodstream infections. Lancet Infect Dis 4:751–760 [View Article][PubMed]
    [Google Scholar]
  18. Sancho-Tello S., Bravo D., Borrás R., Costa E., Muñoz-Cobo B., Navarro D. 2011; Performance of the LightCycler SeptiFast test Mgrade in detecting microbial pathogens in purulent fluids. J Clin Microbiol 49:2988–2991 [View Article][PubMed]
    [Google Scholar]
  19. Stoodley P., Braxton E. E. Jr, Nistico L., Hall-Stoodley L., Johnson S., Quigley M., Post J. C., Ehrlich G. D., Kathju S. 2010; Direct demonstration of Staphylococcus biofilm in an external ventricular drain in a patient with a history of recurrent ventriculoperitoneal shunt failure. Pediatr Neurosurg 46:127–132 [View Article][PubMed]
    [Google Scholar]
  20. Vince A., Lepej S. Z., Barsić B., Dusek D., Mitrović Z., Serventi-Seiwerth R., Labar B. 2008; LightCycler SeptiFast assay as a tool for the rapid diagnosis of sepsis in patients during antimicrobial therapy. J Med Microbiol 57:1306–1307 [View Article][PubMed]
    [Google Scholar]
  21. Yanagihara K., Kitagawa Y., Tomonaga M., Tsukasaki K., Kohno S., Seki M., Sugimoto H., Shimazu T., Tasaki O. et al. other authors 2010; Evaluation of pathogen detection from clinical samples by real-time polymerase chain reaction using a sepsis pathogen DNA detection kit. Crit Care 14:R159 [View Article][PubMed]
    [Google Scholar]
  22. Zubert S., Funk D. J., Kumar A. 2010; Antibiotics in sepsis and septic shock: like everything else in life, timing is everything. Crit Care Med 38:1211–1212 [View Article][PubMed]
    [Google Scholar]
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