1887

Abstract

This study prospectively evaluated an 18S rRNA gene-targeted real-time PCR approach in comparison with standard blood culture (BC) diagnostics for rapid diagnosis of candidaemia in a large study population of 384 patients, including 902 whole blood samples from 468 infectious episodes (IEs) of 329 adults and 55 children with haematological malignancies and various forms of immunodeficiency, and intensive care unit patients. Seven out of eight BC-proven cases (87.5 %) of candidaemia and seven out of twelve BC-positive samples (58.3 %) were positive by the -specific PCR. A positive PCR result was also obtained for 28/460 BC-negative samples from IEs, including 8 patients with culture-confirmed infection at primary sterile body sites. Of the PCR-positive, culture-negative patients, more than 50 % received systemic antifungal therapy. In 432/460 BC-negative IEs, the specific-PCR was negative, resulting in a negative predictive value of 99.8 %. In conclusion, the specific-PCR approach facilitates rapid detection of DNA in blood samples of patients at risk of candidaemia within a few hours. Although standard BC diagnostics appear to remain indispensable for the detection of all cases of candidaemia, this PCR assay allowed the detection of candidaemia at a mean of 3 days earlier than BC diagnostics. Thus, it enables earlier antifungal therapy for patients with suspected candidaemia and may prevent further complications.

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2009-08-01
2024-03-28
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References

  1. Ahmad S., Khan Z., Mustafa A. S., Khan Z. U. 2002; Seminested PCR for diagnosis of candidemia: comparison with culture, antigen detection, and biochemical methods for species identification. J Clin Microbiol 40:2483–2489 [CrossRef]
    [Google Scholar]
  2. Alam F. F., Mustafa A. S., Khan Z. U. 2007; Comparative evaluation of (1, 3)- β -d-glucan, mannan and anti-mannan antibodies, and Candida species-specific snPCR in patients with candidemia. BMC Infect Dis 7:103–111 [CrossRef]
    [Google Scholar]
  3. Bone R. C., Balk R. A., Cerra F. B., Dellinger R. P., Fein A. M., Knaus W. A., Schein R. M., Sibbald W. J. 1992; Definitions for sepsis and organ failure and guidelines for the use of innovative therapies in sepsis. The ACCP/SCCM Consensus Conference Committee. American College of Chest Physicians/Society of Critical Care Medicine. Chest 101:1644–1655 [CrossRef]
    [Google Scholar]
  4. Bretagne S., Costa J. M. 2006; Towards a nucleic acid-based diagnosis in clinical parasitology and mycology. Clin Chim Acta 363:221–228 [CrossRef]
    [Google Scholar]
  5. Dunyach C., Bertout S., Phelipeau C., Drakulovski P., Reynes J., Mallie M. 2008; Detection and identification of Candida spp. in human serum by LightCycler real-time polymerase chain reaction. Diagn Microbiol Infect Dis 60:263–271 [CrossRef]
    [Google Scholar]
  6. Einsele H., Hebart H., Roller G., Loffler J., Rothenhofer I., Muller C. A., Bowden R. A., van Burik J., Engelhard D. other authors 1997; Detection and identification of fungal pathogens in blood by using molecular probes. J Clin Microbiol 35:1353–1360
    [Google Scholar]
  7. Engel C., Brunkhorst F. M., Bone H. G., Brunkhorst R., Gerlach H., Grond S., Gruendling M., Huhle G., Jaschinski U. other authors 2007; Epidemiology of sepsis in Germany: results from a national prospective multicenter study. Intensive Care Med 33:606–618 [CrossRef]
    [Google Scholar]
  8. Gebert S., Siegel D., Wellinghausen N. 2008; Rapid detection of pathogens in blood culture bottles by real-time PCR in conjunction with the pre-analytic tool MolYsis. J Infect 57:307–316 [CrossRef]
    [Google Scholar]
  9. Horz H. P., Scheer S., Huenger F., Vianna M. E., Conrads G. 2008; Selective isolation of bacterial DNA from human clinical specimens. J Microbiol Methods 72:98–102 [CrossRef]
    [Google Scholar]
  10. Innings A., Ullberg M., Johansson A., Rubin C. J., Noreus N., Isaksson M., Herrmann B. 2007; Multiplex real-time PCR targeting the RNase P RNA gene for detection and identification of Candida species in blood. J Clin Microbiol 45:874–880 [CrossRef]
    [Google Scholar]
  11. Klingspor L., Jalal S. 2006; Molecular detection and identification of Candida and Aspergillus spp. from clinical samples using real-time PCR. Clin Microbiol Infect 12:745–753 [CrossRef]
    [Google Scholar]
  12. Kumar A., Roberts D., Wood K. E., Light B., Parrillo J. E., Sharma S., Suppes R., Feinstein D., Zanotti S. other authors 2006; Duration of hypotension before initiation of effective antimicrobial therapy is the critical determinant of survival in human septic shock. Crit Care Med 34:1589–1596 [CrossRef]
    [Google Scholar]
  13. Laupland K. B., Kirkpatrick A. W., Church D. L., Ross T., Gregson D. B. 2004; Intensive-care-unit-acquired bloodstream infections in a regional critically ill population. J Hosp Infect 58:137–145 [CrossRef]
    [Google Scholar]
  14. Maaroufi Y., Heymans C., De Bruyne J. M., Duchateau V., Rodrigues-Villalobos H., Aoun M., Crokaert F. 2003; Rapid detection of Candida albicans in clinical blood samples by using a TaqMan-based PCR assay. J Clin Microbiol 41:3293–3298 [CrossRef]
    [Google Scholar]
  15. Moreira-Oliveira M. S., Mikami Y., Miyaji M., Imai T., Schreiber A. Z., Moretti M. L. 2005; Diagnosis of candidemia by polymerase chain reaction and blood culture: prospective study in a high-risk population and identification of variables associated with development of candidemia. Eur J Clin Microbiol Infect Dis 24:721–726 [CrossRef]
    [Google Scholar]
  16. Tirodker U. H., Nataro J. P., Smith S., Las Casas L., Fairchild K. D. 2003; Detection of fungemia by polymerase chain reaction in critically ill neonates and children. J Perinatol 23:117–122 [CrossRef]
    [Google Scholar]
  17. Wisplinghoff H., Bischoff T., Tallent S. M., Seifert H., Wenzel R. P., Edmond M. B. 2004; Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study. Clin Infect Dis 39:309–317 [CrossRef]
    [Google Scholar]
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